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ERK2 but not ERK1 mediates HGF-induced motility innon-small cell lung carcinoma cell lines

Simone Radtke1,2,*, Mina Milanovic1,*, Carine Rosse1,3, Manu De Rycker1, Sylvie Lachmann1, Andrew Hibbert1,Stephanie Kermorgant2,` and Peter J. Parker1,4,`

1London Research Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK2Centre for Tumour Biology, Barts Cancer Institute, Queen Mary University of London, John Vane Science Centre, Charterhouse Square,London EC1M 6BQ, UK3Membrane and Cytoskeleton Dynamics Laboratory, CNRS, Paris Cedex, France4Division of Cancer Studies, Guy’s Campus, London SE1 1UL, UK

*These authors contributed equally to this work`Authors for correspondence (peter.parker@cancer.org.uk; s.kermorgant@qmul.ac.uk)

Accepted 11 March 2013Journal of Cell Science 126, 2381–2391� 2013. Published by The Company of Biologists Ltddoi: 10.1242/jcs.115832

SummaryAberrant signalling of receptor tyrosine kinases (RTKs), such as c-Met, the receptor for hepatocyte growth factor (HGF), has beenimplicated in the oncogenesis of various tumours including non-small cell lung carcinoma (NSCLC). Through its pro-migratoryproperties, c-Met has been implicated specifically in the process of tumour metastasis, demanding a better understanding of the

underlying signalling pathways. Various players downstream of c-Met have been well characterised, including the extracellular-signal-regulated kinases (ERKs) 1 and 2. In a small interfering RNA (siRNA)-based high-throughput wound healing screen performed in A549lung carcinoma cells, we identified ERK2 but not ERK1 as a strong mediator of HGF-induced motility. This finding was confirmed in

several NSCLC cell lines as well as in HeLa cells. One known substrate for ERK kinases in cell migration, the focal adhesion proteinpaxillin, was also one of the hits identified in the screen. We demonstrate that HGF stimulation results in a time-dependentphosphorylation of paxillin on serine 126, a process that can be blocked by inhibition of the ERK1/2 upstream kinase mitogen-activated

protein kinase/ERK kinase 1 (MEK1) or inhibition of glycogen synthase kinase 3 (GSK3). Further, we show that paxillin turnover atfocal adhesions is increased upon stimulation by HGF, an effect that is dependent on serine residues 126 (GSK3 site) and 130 (ERK site)within paxillin. In line with the isoform-specific requirement of ERK2 for HGF-mediated migration in lung tumour cell models, ERK2

but not ERK1 is shown to be responsible for paxillin serine 126 phosphorylation and its increased turnover at focal adhesions.

Key words: HGF, c-Met, Motility, Wound healing, High-throughput screen, Signalling, siRNA, ERK2, Paxillin

IntroductionCell migration is a fundamental process important for normal

tissue homeostasis and is triggered by various environmentalstimuli and their downstream signalling pathways, ultimately

resulting in the rearrangement of the cytoskeleton, reorganisation

of the microtubule organising centre, extension of membrane

protrusions, adhesion to the substratum and generation of forcesthat physically move a cell (reviewed by Vicente-Manzanares

et al., 2005). Aberrations in components of these signal

transduction pathways can lead to tumour dissemination and

invasion to surrounding tissues (reviewed by Friedl and Wolf,2003), playing a major role in cancer development and

progression.

Hepatocyte growth factor (HGF) can trigger cell migration

through binding and activation of its only known receptor, c-Met, a

receptor tyrosine kinase. The activated receptor serves as a

docking site for various adaptor and signalling proteins, leading tothe disassembly of adherens junctions, increased cell motility,

survival and branching morphogenesis (Potempa and Ridley,

1998; Bowers et al., 2000; Zhang et al., 2002; Kermorgant and

Parker, 2008). A role of c-Met has been observed in many humanmalignancies, including gastric and oesophageal carcinoma,

medulloblastoma and Non Small Cell Lung Cancer (NSCLC)

(Tong et al., 2004; Gao and Vande Woude, 2005; Miller et al.,

2006). Importantly, HGF/c-Met are believed to play a major role in

the progression to tumour metastasis (reviewed by Peruzzi and

Bottaro, 2006; Sattler and Salgia, 2007) so that a detailed

understanding of processes underlying c-Met-dependent cancer

cell migration is important for providing opportunities for the

development of new treatments for these diseases. Several

downstream pathways of c-Met have been well characterised

(reviewed by Trusolino et al., 2010); thus the importance of the

ERK1/2 pathway in HGF-mediated cellular motility has been

demonstrated in various cellular models (Tanimura et al., 1998;

Zeigler et al., 1999; Karihaloo et al., 2001; Tanimura et al., 2002;

Kermorgant et al., 2004; Menakongka and Suthiphongchai, 2010).

As c-Met overexpression, mutation or increased activation has

been observed in NSCLC (reviewed by Sattler et al., 2011), we

used the lung carcinoma cell line A549 to perform an siRNA-

based wound healing screen in order to study the signalling

pathways important for c-Met mediated migration and identified

ERK2 but not ERK1 as one of the strongest hits. So far, the

specific contribution of ERK1 and ERK2 has not been studied

with respect to the HGF-signalling cascade. Here we show that

HGF-induced motility is specifically mediated by ERK2 in

several cell lines, and this finding correlated with the observation

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that ERK2 but not ERK1 was required for the HGF-induced

phosphorylation of the focal adhesion scaffold protein paxillin on

serine 126, a site which has been described to be phosphorylated

by glycogen synthase kinase (GSK) 3 upon priming of the

adjacent serine 130 by ERKs (Cai et al., 2006). The important

role of this phosphorylation in HGF-mediated migration was

demonstrated by the finding that the siRNA-mediated depletion

of endogenous paxillin results in a decrease in HGF-mediated

migration, an effect which can be rescued by wild type paxillin

but not a mutant in which serine 126 and the adjacent serine 130

are mutated to alanine. In line with the motility data, we can

show that HGF stimulation results in an increased disassembly of

paxillin-containing focal adhesions, an effect which can be

inhibited by knockdown of ERK2 but not ERK1. Thus we

propose that paxillin is an important mediator downstream of

ERK2 in HGF-induced motility.

ResultsIdentification of ERK2 in an siRNA-based wound healing

screen

We set up an siRNA-based high throughput wound healing assay

using the A549 lung carcinoma cell model. Altogether, 1115

siRNA pools targeting the human kinome, a set of motor proteins

as well as genes implicated in motility were transfected and a

non-targeting control (NTC) siRNA and an siRNA against c-Met

served as negative and positive controls respectively. An siRNA

targeting the tyrosine kinase receptor Macrophage-stimulating

protein receptor (MST1R), a c-Met paralog (Ronsin et al., 1993;

Gaudino et al., 1994), was included as an additional control. A

detailed schematic overview of how the Primary screen was

carried out is depicted in supplementary material Fig. S1.

Cells transfected with non-targeting siRNA displayed roughly

a twofold increase in motility speed upon HGF treatment, an

effect which was almost completely abrogated upon transfection

of the c-Met siRNA. In contrast, knockdown of the related

receptor MST1R had a significant effect both on the basal and

HGF-induced motility speed, retaining a twofold increase

between the two wound healing speeds (Fig. 1A). To have a

read out for the effect of the transfected siRNAs on the HGF-

response, an HGF-induction score was calculated as exemplified

in supplementary material Fig. S2. The normalised induction

scores obtained for the negative and positive controls used are

depicted in Fig. 1B. As expected, c-Met knockdown resulted in a

strong decrease of the induction score to a value close to 0,

Fig. 1. High-throughput wound healing screen. A549 cells were transfected with 1115 Dharmacon siRNA pools directed against different genes. As a negative

control a non-targeting siRNA was used (NTC), siRNAs targeting the HGF-receptor cMet (MET) and the related tyrosine kinase receptor Mst1R were used as

additional controls. Wound healing assays were carried out 72 hours post transfection. Raw migration speeds were calculated for control treated and HGF-treated

cells and using the two sets of raw migration data, HGF-induction scores were calculated as described in Material and Methods. (A,B) Negative and positive

controls used in the screens. Raw migration speeds were calculated for control treated and HGF-treated cells. Averages and standard deviations calculated from all

control values (A) and HGF-induction scores (B) obtained during the screen are depicted. (C) HGF-induction scores obtained during the primary screen. The

obtained HGF-induction scores for all transfected siRNAs are shown in ascending order. Black dotted lines represent the thresholds used for classifying siRNAs

that significantly increase or decrease the HGF-response. The black circle and the black triangle represent the values obtained for MET and ERK2, respectively,

that were part of the assayed kinome library. (D) Knockdown efficiencies of transfected siRNAs. A549 cells were transfected with the siRNA pools as indicated

and harvested 72 hours post transfection. Total cellular lysates were generated and knockdown efficiency was monitored by western blot analysis using the

antibodies indicated. Representative blots are depicted. (E,F) siRNAs targeting MET and ERK2 strongly impair HGF-induced wound healing. A549 cells were

transfected with the siRNA pools as indicated and 72 hours post transfection, wound healing assays were carried out. Average wound healing speeds were

calculated for control-treated and HGF-treated cells (E). The HGF wound healing score was calculated from both values. Means 6 s.e.m. derived from three

independent experiments are depicted. Statistical significance was determined using paired Student’s t-test (F); *P,0.05, ***P,0.001.

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whereas knockdown of MST1R only slightly affected the

induction score. Fig. 1C displays the induction scores obtained

during the primary screen in an ascending order. Thresholds

were chosen as described in Materials and Methods. While 85

siRNAs were classified as negative regulators of the HGF-

response, 43 siRNAs resulted in an increased HGF-response.

Data obtained for all siRNAs is provided in supplementary

material Table S1. MET kinase itself (black circle) and ERK2

(black triangle), which were both part of the library targeting the

human kinome, came up among the top 20 hits, serving as a

proof of principle that the screen was able to identify players

involved in HGF-mediated signalling. These results could be

confirmed in further experiments, where for both siRNA pools a

good knockdown was observed by western blot (Fig. 1D). In

wound healing experiments, ERK2 but not MET depletion

resulted in a strong decrease of the basal wound healing speed

(Fig. 1E). As expected, MET knockdown resulted in an almost

completely abrogated HGF-induced migration, while ERK2

depletion reduced the HGF-induced migration by more than

50% (Fig. 1F).

Confirmation of the migratory phenotype in ERK2-depleted

cells

In the primary screen, we used a wound healing assay to find

players involved in HGF-induced cell migration. Not surprisingly

given its role in the regulation of cell proliferation, we observed a

strong effect of ERK2 depletion on cell viability (Fig. 2A). To

confirm that the results obtained using the wound healing assays

were not merely a consequence of reduced cell proliferation, we

carried out live cell imaging followed by cell tracking to monitor

single cell motility speed after siRNA knockdown. In Fig. 2B

representative cell tracks in wounds of cells transfected with non-

targeting control siRNA (NTC), MET siRNA or ERK2 siRNA

are depicted. In NTC-transfected cells, HGF incubation led to

approximately a twofold increase in the single cell motility

speed, an effect which was almost completely abrogated by MET

knockdown and also strongly inhibited (75%) by knockdown of

ERK2 (Fig. 2C). Similar results were achieved using Transwell

assays as a distinct migratory model (Fig. 2D), thus confirming

the migratory phenotype observed in the wound healing assays.

Knockdown of ERK2 but not ERK1 has an effect on HGF-

induced motility

While ERK2 was one of the strongest hits identified in this

screen, intriguingly ERK1, the second ERK isoform, was not

identified as a hit. To assess the validity of this finding, we

compared the effect of ERK2 and ERK1 knockdown side by side.

Silencing of both ERK2 and ERK1 resulted in an efficient

knockdown as evidenced by western blot analysis (Fig. 3A) and

depletion of both ERK isoforms had a very similar effect on cell

viability (Fig. 3B). Both ERK1 and ERK2 depletion resulted in a

significantly reduced basal motility with a consistently greater

effect of ERK1 knockdown (Fig. 3C, light grey bars). However

by contrast, ERK2 knockdown alone strongly impaired the HGF

response, while ERK1 silencing did not, with HGF still able to

induce a strong increase in wound healing speed (Fig. 3C, right

panel). Interestingly, the MEK1 inhibitor U0126 gave similar

results as the single knockdown of ERK2 (Fig. 3D). The specific

role of ERK2 in HGF-mediated motility was not restricted to

A549 cells, since also in CALU-1 and SKMES-1 lung carcinoma

cell lines as well as in HeLa cells, HGF-induced cell motility was

affected by knockdown of ERK2 but not ERK1 (Fig. 3E).

Paxillin is required for HGF-mediated migration

Various substrates of the ERK kinases have been reported to play

a role in cellular migration, including the focal adhesion protein

paxillin, which has been shown previously to be important for

HGF-induced migration (Liu et al., 2002; Ishibe et al., 2003;

Ishibe et al., 2004). Interestingly, paxillin was another strong hit

in the wound healing screen we performed and indeed this data

could be confirmed in cell tracking experiments both in A549 and

in HeLa cells (Fig. 4A). Knockdown of paxillin did not affect

Fig. 2. Validation of the migratory phenotype. (A) Knockdown of ERK2

significantly reduces cell viability. Cell viability of transfected cells was

assayed using CellTiter-glo as described in Materials and Methods. Values

were normalised to data obtained from NTC transfected cells.

Means 6 s.e.m. derived from four independent experiments are depicted.

(B,C) Live cell imaging and single cell tracking. Transfected A549 cells were

grown to confluency and 72 hours post transfection, wounding was

performed, cells were incubated in medium containing 0.5% FCS with or

without HGF and live cell imaging was carried out for 12 hours. From the

resulting pictures, movies were generated and at least 20 cells per movie were

tracked and cell tracks analysed as described in Materials and Methods. For

each condition, median single cell motility speeds were calculated from at

least 40 cell tracks per experiment. Representative tracks from one

experiment are depicted (B). Average single cell speed values and the

resulting HGF migration scores obtained from four independent experiments

are depicted (C). (D) Transwell assays. 72 hours post transfection, cells were

trypsinised and plated onto 24-well transwell dishes or normal 24-well tissue

culture plates in duplicate. After incubation for 3 hours with or without HGF,

the cell numbers of plated cells and of cells that had migrated through the

transwells were estimated using CellTiter-glo and the percentage of migrated

cells was calculated. The average value obtained from NTC transfected, HGF-

treated cells was set to 1. Means 6 s.e.m. obtained from three independent

experiments are depicted in the left diagram; the resulting average HGF

migration scores are depicted in the right diagram. In all cases, statistical

significance was determined by paired Student’s t-test; *P,0.05, **P,0.01,

***P,0.001; ns, not significant.

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HGF-induced MET or ERK phosphorylation in A549 cells

(Fig. 4B) or HeLa cells (data not shown). To further investigate

the role of paxillin in HGF-mediated motility, we monitored

paxillin turnover in HeLa cells by total internal reflection

fluorescence (TIRF) microscopy. When analysing focal

adhesions containing GFP-tagged paxillin over time as

described (Berginski et al., 2011), we saw an increase of focal

adhesion disassembly rates upon HGF stimulation (Fig. 4C).

Paxillin turnover has been shown to be regulated by

phosphorylation (Webb et al., 2004; Berginski et al., 2011) and

indeed, HGF stimulation resulted in an increase in the

phosphorylation of serine 126 in paxillin, an effect which was

maximal after about 2 hours of HGF stimulation (Fig. 4D).

Serine 126 has been described to be phosphorylated by glycogen

synthase kinase 3 (GSK3) upon priming of the adjacent serine

130 by ERK (Cai et al., 2006). Consistent with this, HGF-

mediated phosphorylation of serine 126 was abrogated both by

incubation with the MEK1 inhibitor U0126 or the GSK3 inhibitor

IX (Fig. 4E). To further investigate the role of these two

phosphorylation sites in HGF-mediated migration, we

constructed GFP-tagged paxillin where both serine 126 and

serine 130 were mutated to alanine (GFP-PaxAA). These

constructs were transfected into HeLa cells and adhesion

turnover was monitored by TIRF microscopy. As seen before,

HGF resulted in an increase of the disassembly rate of focal

adhesions, an effect which was strongly inhibited in focal

adhesions containing the paxillin mutant (Fig. 4F). Furthermore,

in HeLa cells, siRNA-mediated knockdown of endogenous

paxillin by a single oligo resulted in a significantly reduced

HGF-mediated migration as monitored by live cell imaging, an

effect which could be rescued by overexpression of GFP-tagged

wild-type (wt) paxillin but not the mutant GFP-PaxAA (Fig. 4G).

ERK2 knockdown results in diminished HGF-induced

paxillin phosphorylation and turnover

Given the important role of paxillin phosphorylation in HGF-

mediated migration, we next monitored phosphorylation of

paxillin upon knockdown of either ERK1 or ERK2.

Fig. 3. ERK2 but not ERK1 is essential for HGF-induced wound healing. (A) Knockdown efficiency of siRNAs used. A549 cells were transfected with the

siRNA pools indicated and 72 hours post transfection total cellular lysates were generated and expression of ERK1 and ERK2 was monitored by western blot

analysis. A representative blot is depicted in the left panel. Blots were quantified as described in Materials and Methods and the relative expression was calculated.

Means 6 s.e.m. derived from three independent experiments are displayed. (B) ERK2 and ERK1 knockdown have a similar effect on cell viability. A549 cells

were transfected with the indicated siRNAs and harvested 72 hours post transfection. Cell viability was assayed as described in Materials and Methods.

Means 6 s.e.m. from three independent experiments are depicted. (C) ERK2 but not ERK1 knockdown affects HGF-induced motility. A549 cells were

transfected with the siRNAs indicated and 72 hours post transfection, cell wounding and live cell imaging were performed. Cell tracking was carried out and

single cell motility speeds (left panel) as well as HGF migration scores (right panel) were calculated as described in Materials and Methods. Means 6 s.e.m.

derived from three independent experiments are depicted. (D) Effect of MEK1 inhibition. A549 cells were treated with DMSO or U0216 (U0) for 20 minutes

before wounding was carried out. Cell motility was monitored by videomicroscopy. Cell tracking was carried out and HGF migration scores were calculated from

the obtained single cell motility speeds as described in Materials and Methods. Average normalised single cell motility speeds as well as HGF migration scores are

depicted. To determine statistical significance, paired Student’s t-test was used. (E) ERK2 is specifically needed for HGF-induced wound healing in various cell

lines. The indicated siRNAs were transfected into HeLa, CALU-1 and SKMES-1 cells. At 72 hours post transfection, cells were wounded and wound healing was

monitored using videomicroscopy. Cell tracking was carried out and HGF migration scores were calculated from the obtained single cell motility speeds as

described in Materials and Methods. Average HGF-migration scores are depicted. In all cases, statistical significance was determined using paired Student’s t-test;

*P,0.05, **P,0.01.

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Interestingly, in cells transfected with siRNA against ERK1, HGF

was still able to cause an increase in paxillin phosphorylation,

however, this effect was strongly inhibited when ERK2 was

depleted by siRNA (Fig. 5A). Experiments using the deconvoluted

ERK2 siRNA pool corroborated these findings (supplementary

material Fig. S3). Furthermore, this effect was not restricted to

A549 cells, as also in CALU, SKMES and HeLa cells, ERK2

knockdown led to a strong decrease in HGF-mediated paxillin

phosphorylation (supplementary material Fig. S4).

It has been shown that the phosphatidylinositide 3-kinase

(PI3K)/AKT-mediated serine phosphorylation of GSK3

negatively regulates its activity (Wang et al., 1994; Cross et al.,

1995). Indeed we saw an increase of AKT and GSK3

phosphorylation upon HGF treatment (supplementary material

Fig. S5), and GSK3 phosphorylation could be almost completely

prevented by preincubation with the PI3K inhibitor LY294002

(supplementary material Fig. S5B). To exclude the possibility

that the effect of ERK2 knockdown on paxillin phosphorylation

Fig. 4. Paxillin is required for HGF-induced migration and is phosphorylated upon HGF treatment. (A) Paxillin depletion results in reduced cell motility

upon HGF treatment. A549 cells and HeLa cells were transfected with the siRNAs indicated and 48 hours post transfection cell wounding and live cell imaging

was performed. Cell tracking was carried out and single cell motility speeds as well as HGF migration scores were calculated as described in Materials and

Methods. Average normalised single cell motility speeds are shown in the left panel; HGF migration scores are shown in the right panel. Means 6 s.e.m. derived

from four independent experiments are depicted. Statistical significance was determined using paired Student’s t-test. (B) Paxillin depletion does not affect the

HGF-induced phosphorylation of MET and ERK1/2. A549 cells were transfected with control siRNA (NTC) or siRNA targeting paxillin. At 48 hours post

transfection, cells were treated for 2 hours with HGF or left untreated, total cellular lysates were generated and analysed by western blot as indicated.

Representative blots are shown. (C) HGF treatment increases the paxillin turnover in HeLa cells. HeLa cells transiently expressing GFP-tagged paxillin were

cultured in 10% FCS or 10% FCS supplemented with HGF and monitored over 30 minutes using TIRF microscopy. The resulting movies were used to calculate

the assembly and disassembly rates of paxillin containing focal adhesions, as described in the Materials and Methods. At least 10 movies per condition were

analysed. Statistical significance was determined using the unpaired Student’s t-test. (D,E) HGF-mediated paxillin phosphorylation on serine 126 is ERK- and

GSK3-dependent. Cells were treated with HGF for the indicated time points and total cellular lysates were generated (D). For inhibitor experiments, cells were

pretreated with DMSO, U0216 (U0) or GSK3 inhibitor IX (IX) for 20 minutes and then treated with HGF or left untreated for 2 hours and total cellular lysates

generated (E). Lysates were analysed by western blot with the antibodies indicated. Representative blots are shown in the left panel. Blots were quantified using

ImageJ software and values normalised to those obtained for tubulin. Average normalised values from three independent experiments are depicted. (F) Altered

HGF-induced disassembly rates in cells expressing GFP-paxillin-AA. HeLa cells transiently expressing GFP-tagged wild-type paxillin (wt) or mutated paxillin

(AA) were cultured in 10% FCS or 10% FCS supplemented with 50 ng/ml HGF and monitored over 30 minutes using TIRF microscopy. The resulting movies

were used to calculate the assembly and disassembly rates of paxillin-containing focal adhesions, as described in the Materials and Methods. At least 10 movies

per condition were analysed. (G) GFP-paxillin-wt but not GFP-paxillin-AA can rescue the HGF-induced migration in HeLa cells depleted of endogenous paxillin.

HeLa cells were transfected with non-targeting control siRNA or an siRNA oligo targeting paxillin and 24 hours post transfection were further transfected with a

cDNA for GFP, GFP-paxillin-wt or GFP-paxillin-AA. The next day, a wound healing assay was performed and cell migration was monitored by live cell imaging

and single cell migration speeds determined by cell tracking. For each condition, six movies were analysed. Average single cell migration speeds are depicted.

Statistical significance was determined using one-way Anova; *P,0.05, ***P,0.001; ns, not significant.

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could be due to altered GSK3 activity, we therefore also

monitored GSK3 phosphorylation, however, there was no

difference in the HGF-mediated GSK3 phosphorylation upon

ERK2 knockdown (Fig. 5A).

Finally, when monitoring the effect of ERK1 and ERK2

knockdown on paxillin turnover in HGF treated HeLa cells, we

saw that assembly rates were not affected by either ERK1 or

ERK2 knockdown, whereas ERK2 but not ERK1 knockdown

resulted in a significant decrease in the rate of paxillin

disassembly (Fig. 5B). We conclude that HGF via ERK2

induces phosphorylation of paxillin, thus mediating an increase

both in paxillin turnover at focal adhesions and cell migration.

Overexpression of ERK2 but not ERK1 can efficientlyrescue the effects of siRNA-mediated knockdown of

endogenous ERK2

The fact that knockdown of ERK2 but not ERK1 resulted in loss

of HGF-induced migration and paxillin phosphorylation could

theoretically be explained by the fact that ERK2 is the prominent

isoform expressed. We therefore tested whether the effect of the

knockdown of endogenous ERK2 could be rescued by either

overexpression of ERK1 or ERK2. A549 cells were transfected

with a single siRNA oligonucleotide targeting ERK2 (E2-1), and

24 hours later, cDNA for either HA-tagged ERK1 or ERK2 was

transfected; an empty vector served as a negative control. Paxillin

phosphorylation and ERK activation were monitored by western

blotting. Both ERKs were expressed to a similar extent (Fig. 5A,

left panel), and both ERK1 expression and ERK2 expression

could rescue overall ERK-activation to a similar extent (Fig. 5A,

upper graph). However, only ERK2 but not ERK1

overexpression could rescue the phosphorylation of paxillin

(Fig. 5A, lower graph). Similarly, ERK2 and not ERK1

expression resulted in the rescue of HGF-induced single cell

motility (Fig. 5B). To assess these findings in non-lung tumour

cells, we also carried out rescue experiments in HeLa cells. In

these experiments ERK1 was much more effectively expressed

than ERK2, and while both ERK isoforms were able to rescue the

HGF-mediated paxillin phosphorylation to some extent, ERK2

was more efficient than ERK1 in doing so, such that there is an

almost twofold relative efficiency of activated ERK2 compared

to activated ERK1 in mediating the HGF-induced paxillin

phosphorylation (supplementary material Fig. S6A). In line

with these findings, when monitoring the ability of ERK1 and

ERK2 to rescue HGF-mediated cell motility, we similarly noted a

greater ability of ERK2 than ERK1 to do so (supplementary

material Fig. S6B). A schematic model summarising our findings

is displayed in Fig. 6C. We conclude that HGF can regulate

paxillin phosphorylation via its effects on GSK3 and ERK2. Our

finding that ERK2 is the dominant ERK isoform in mediating

paxillin phosphorylation can explain the strong impact of ERK2

but not ERK1 knockdown on Paxillin turnover and cell migration.

DiscussionUnderstanding the detailed signalling pathways required for

HGF-mediated migration is of great interest for better

understanding and therefore selectively targeting aberrant

signalling of c-Met in cancer metastasis. In performing an

siRNA based high throughput wound healing screen in the

NSCLC cell line A549, one of the strongest hits identified was

ERK2, inhibiting the HGF-mediated migration consistently by

more than 50%. ERK1 and 2 have been implicated in migration

in various cell types, in response to different matrix proteins,

various growth factors and other stimuli (Karihaloo et al., 2001;

Huang et al., 2004). Therefore, the finding that ERK2 is one of

Fig. 5. ERK2 knockdown inhibits HGF-mediated paxillin phosphorylation and decreases paxillin turnover. (A) ERK2 knockdown prevents HGF-induced

paxillin phosphorylation. A549 cells were transfected with control siRNA (NTC) or siRNA targeting ERK2 or ERK1. At 48 hours post transfection, cells were

treated for 2 hours with HGF or left untreated, total cellular lysates were generated and analysed by western blot as indicated. Representative blots are shown on

the left panel. Blots were quantified using ImageJ software and values normalised to those obtained for tubulin. Average normalised values are depicted. To

determine statistical significance, two-way Anova was carried out, followed by Bonferroni post hoc test. (B) ERK2 knockdown decreases the paxillin disassembly

rate in HGF-treated HeLa cells. HeLa cells depleted of ERK1 or ERK2 by siRNA for 48 hours and transiently expressing GFP-tagged paxillin were cultured in

10% FCS supplemented with HGF and monitored over 30 minutes using TIRF microscopy. The resulting movies were used to calculate the assembly and

disassembly rates of paxillin containing focal adhesions, as described in the Materials and Methods. At least 10 movies per condition were analysed. Statistical

significance was determined using one-way Anova followed by Bonferroni post hoc test; ***P,0.001; ns, not significant.

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the strongest hits isolated from the screen provides a proof-of-

principle in terms of screen validity. However it is striking that in

the screen, only ERK2 but not ERK1 was found to affect HGF-

induced migration. While a role for the ERK1/2 pathway in HGF-

mediated motility has been shown before, most studies related to

ERK1/2 signalling have made no functional distinction between

the two isoforms and whether ERK1 and ERK2 play similar or

distinct roles in HGF dependent motility has remained unknown.

The evidence here demonstrates that it is ERK2 and not ERK1

that plays a role downstream of MET in lung tumour models and

that there is a strong bias towards ERK2 in other cell types tested.

Until recently isoform specificity of ERKs has not been widely

addressed. Indeed several studies have shown a redundant

function of ERK1 and ERK2 in cell proliferation (Srinivasan

et al., 2009; Voisin et al., 2010). Likewise, in a mouse model of

K-Ras induced NSCLC, elimination of individual ERK kinases

resulted in a similar small decrease of K-Ras induced

oncogenesis, while only elimination of both ERK1 and ERK2

impaired tumour development (Blasco et al., 2011). In line with

these publications, when monitoring cell viability, we also find a

similar effect upon knockdown of either ERK isoform. In

contrast to such a redundant function of both ERK isoforms, there

are recent studies that have identified some isoform specific

functions. Thus, ERK1 and ERK2 have been demonstrated to

fulfil different roles in Ras-dependent signalling (Vantaggiato

et al., 2006) and long-term survival of hepatocytes (Fremin et al.,

2009). While some differences in ERK isoform signalling have

been attributed to differences in the N-terminal domain of both

isoforms (Marchi et al., 2008), other studies have shown that

differences might be due to the expression level of each isoform

(Lefloch et al., 2008; Lefloch et al., 2009). Indeed ERK2 is

expressed to a much higher level than ERK1 in all of the cell

lines we examined, although some properties of these cells such

as basal migration rates were more sensitive to ERK1 knock-

down than to ERK2 knockdown. Notwithstanding this

preferential effect of ERK1, to examine whether our findings

are due to the fact that ERK2 is the prominent isoform, we also

carried out rescue experiments using HA-tagged ERK constructs.

In A549 cells, only ERK2 but not ERK1 could rescue the

knockdown of endogenous ERK2. Similarly, while both ERK

isoforms were able to rescue the knockdown of ERK2 in HeLa

cells, ERK2 was much more efficient in doing so.

Interestingly it has recently been found that ERK2 but

not ERK1 overexpression induced epithelial-to-mesenchymal

transformation in MCF-10A cells (Shin et al., 2010). Likewise, in

MDA-MB-231 cells, ERK2 but not ERK1 knockdown inhibits

cell invasiveness (von Thun et al., 2012). Consistent with this,

our results suggest that also in case of c-Met driven tumour

metastasis, ERK2 might be the dominant ERK isoform.

Many cellular substrates have been reported to be

phosphorylated by ERKs, including both nuclear and

cytoplasmic proteins. One of the known ERK substrates shown

Fig. 6. ERK2 but not ERK1 overexpression can rescue the effect of siRNA-mediated knockdown of endogenous ERK2. A549 cells were transfected with

control siRNA (NTC) or a single siRNA oligonucleotide targeting ERK2 (E2-1). 24 hours post transfection, cDNAs coding for HA-tagged ERK1 or ERK2 or

empty vector were transfected. Experiments were carried out the next day. (A) HGF-mediated paxillin phosphorylation on serine 126 can be rescued by ERK2.

Cells were treated with HGF for 2 hours and total cellular lysates were generated. Lysates were analysed by western blot with the antibodies indicated.

Representative blots are shown in the left panel. Blots were quantified using ImageJ software and values were normalised to values obtained for tubulin. Average

normalised values derived from five independent experiments are depicted. To determine statistical significance, two-way Anova was carried out, followed by

Bonferroni post hoc test. (B) ERK2 but not ERK1 overexpression can rescue the HGF-induced increase in single cell motility. Cell wounding, live cell imaging

and cell tracking were carried out and single cell motility speeds as well as HGF migration scores were calculated as described in Materials and Methods. Average

normalised single cell motility speeds are shown in the upper panel, HGF migration scores are shown in the lower panel. Means 6 s.e.m. derived from four

independent experiments are depicted. Statistical significance was determined using the paired Student’s t-test. (C) Schematic overview of the cMet–paxillin

signaling axis. The dominant players involved in regulation of paxillin phosphorylation upon HGF are displayed. **P,0.01, ***P,0.001; ns, not significant.

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to play a role in cellular motility is paxillin (reviewed by Brownand Turner, 2004) and paxillin has been implicated in cell

spreading and morphogenesis upon HGF treatment (Liu et al.,2002; Ishibe et al., 2003; Ishibe et al., 2004). Of note, paxillin wasanother strong hit identified in our screen, and we could confirm arole for paxillin in HGF-mediated motility in cell tracking

experiments both in A549 and HeLa cells (Fig. 4A). Variousphosphorylation sites have been identified in paxillin (Webb et al.,2005) and we find an HGF-induced phosphorylation of paxillin on

serine 126. The HGF-mediated phosphorylation of paxillin onTyrosine 118 and serine 83 has been demonstrated before (Liuet al., 2002; Ishibe et al., 2003; Ishibe et al., 2004). However, the

role of serine 126 in HGF-induced motility has not been studied sofar. Of note paxillin phosphorylation at this site is maximal ataround two hours of HGF stimulation and therefore peaks slightlylater than P-MET and P-ERK, a finding we also could reproduce in

other cell lines (data not shown). This small delay can probably beexplained by the fact that serine 126 is not a direct substrate ofERK but in fact phosphorylation of this site has been attributed to

the kinase GSK3, which can only efficiently phosphorylate thissite upon priming phosphorylation of the upstream serine 130 byERK (Cai et al., 2006). Using inhibitors against MEK1 and GSK3,

we can confirm these findings in A549 cells.

GSK3 activity has been shown to be regulated by serine andtyrosine phosphorylation (Wang et al., 1994). Thus phosphoinositide

3-kinases (PI3-K)/AKT-mediated phosphorylation of serine 9inhibits the activity of GSK3b (Cross et al., 1995), whereasphosphorylation of tyrosine 216 positively affects its activity(Wang et al., 1994). HGF is a strong inducer of AKT activity

(reviewed by Trusolino et al., 2010), therefore not surprisingly,we find an increase in phosphorylation of the inhibitory serine9 in GSK3b peaking at 1 hour of HGF stimulation. There is

also a small but reproducible increase in phosphorylation oftyrosine 216, which plateaus between 1 and 4 hours of HGFstimulation (supplementary material Fig. S5). The transient

phosphorylation of serine 9 can explain the delayed kinetics ofpaxillin Serine 126 phosphorylation upon HGF stimulation;indeed when AKT-mediated GSK3 phosphorylation isprevented by the PI3K inhibitor LY294002, we find a shift

in the kinetics to an earlier timepoint (supplementary materialFig. S5B), thus HGF-mediated paxillin phosphorylation seemsto be tightly regulated via the combined outputs of the AKT

and ERK2 signaling pathways.

We show that phosphorylation of serine 126 can be blocked byknockdown of ERK2 but not ERK1 (Fig. 5A), a finding which we

can reproduce in several cell lines (supplementary material Fig.S4). Of note, in Calu as well as in HeLa cells, ERK1 knockdownresults in a small reduction in paxillin phosphorylation, however,

the effect of ERK2 knockdown is much more pronounced.Consistent with these findings, ERK2 is significantly moreefficient than ERK1 in rescuing the diminished paxillinphosphorylation seen in cells transfected with siRNA targeting

ERK2 (Fig. 6A; supplementary material Fig. S6), thus correlatingvery well with the motility data. Indeed, the relevance ofphosphorylation of serine 126 and adjacent serine 130 during

cell migration was demonstrated by the fact that the decreasedHGF-induced motility seen upon paxillin knockdown can berescued by wild-type paxillin but not by a mutant where the GSK3

and ERK sites have been mutated to alanine (Fig. 4G).

We speculate that the ERK2-specific regulation of HGF-induced motility is due in part to specific ERK2-mediated

paxillin phosphorylation, which would in turn affect focal

adhesion dynamics as demonstrated in a different motility

model (Boeckeler et al., 2010). Indeed we find that HGF-

treatment results in an increase of the disassembly rate of

paxillin-containing focal adhesions (Fig. 4C). This effect is

blocked in a mutant paxillin in which serine 126 and serine 130

have been mutated to alanine and hence can no longer be

phosphorylated at these sites (Fig. 4F), and ERK2 but not ERK1

knockdown significantly decreases the disassembly rate of

adhesions in HGF treated A549 cells (Fig. 5B).

Aberrant c-Met signalling has been implicated in the

oncogenesis of NSCLC (reviewed by Cipriani et al., 2009),

where c-Met overexpression correlates with metastatic tendency

and poor prognosis and is found in a high proportion of NSCLCs

as demonstrated by immunohistochemistry (Ma et al., 2005; Ma

et al., 2008). In 5% of NSCLC, c-Met overexpression is due to

gene amplification (Cappuzzo et al., 2009) and, interestingly, c-

Met amplification has been associated with resistance of NSCLC

to gefitinib, an EGFR targeted therapy (Cappuzzo et al., 2009).

Interestingly, paxillin has also recently been found to be

upregulated in NSCLC, correlating with a poor prognosis

(Jagadeeswaran et al., 2008; Wu et al., 2010; Mackinnon et al.,

2011) and it has been reported that there is a high correlation of

paxillin and MET gene copy numbers in NSCLC (Jagadeeswaran

et al., 2008). Therefore, a better understanding of the c-Met-paxillin

signalling axis in HGF-mediated motility is important for

targeting this pathway in NSCLC and our findings that

specifically ERK2 but not ERK1 is required in this pathway

may have important implications for helping to design and

apply specific therapies in future.

Materials and MethodsCell culture

Cells were obtained from Cancer Research UK London Research Institute’s CellServices and cultured at 37 C in a humidified 10% CO2 atmosphere. Cells werecultivated in GIBCOH GlutaMAXTM media (Invitrogen) supplemented with 10%Certified FBS (Invitrogen) and 50 mg Penicillin-Streptomycin (Invitrogen).

HGF was purchased from R&D Systems and used routinely at 10 ng/ml, withthe exception of HeLa cells, which were treated with 40 ng/ml. U0126 was fromCell Signaling and used at 10 mM. GSK3 inhibitor IX was purchased from SantaCruz Biotechnology and used at 5 mM. LY294002 was from Sigma-Aldrich andused at 10 mM.

siRNA libraries

siRNA libraries used in the primary screen were obtained from Dharmacon. Thecomplete list of genes targeted, siRNAs used and Entrez IDs can be found insupplementary material Table S1.

siRNA and cDNA transfections

For the primary screen, each siRNA was transfected in six replicate wells on 96-well plates with Hiperfect transfection reagent (Qiagen), using a reversetransfection protocol according to the manufacturer’s instructions. 20 mltransfection mixture containing both the siRNA and 0.5 ml hiperfect diluted inOptimem were mixed with 80 ml cell suspension to give a final concentration of10 nM siRNA and 86103 cells per well. Growth medium was replaced 24 hourslater, and cells were incubated for a further 48 hours. Unless stated otherwise,experiments were carried out 72 hours post transfection. For some experiments,cells were transfected on 24-well, 12-well or 6-well plates, using the equivalentcell densities, amount of Hiperfect per ml medium and final siRNA concentrationsas for the transfection of 96-well plates. SK MES-1 cells were transfected usingHiperfect or RiboJuice (Novagen) according to manufacturer’s instructions. HeLacells were transfected using lullaby (OZ biosciences). For transfections of cDNAinto A549 and HeLa cells, the LTX reagent (Invitrogen) was used according to themanufacturer’s instructions.

Generation of cDNA constructs

The C-terminal HA tag as well as NheI and XbaI restriction sites were inserted intomouse ERK2 and human ERK1 cDNA respectively by standard PCR techniques

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using appropriate oligonucleotides. The PCR products were inserted into thepcDNA3 vector (Invitrogen) using the in-FusionH HD Cloning Kit (Clontech). TheGFP-paxillin AA mutant was generated using the QuikChange Site-directedMutagenesis kit (Stratagene) with chicken GFP-paxillin as a template, and twooligos containing the double mutation Ser126Ala/Ser130Ala. The validity of allresulting constructs was confirmed by sequencing.

High-throughput wound healing assay

Wound healing assays were carried out on 96-well clear bottom plates 72 hourspost transfection. In order to minimise edge effects, only the 48 central wells ofeach 96-well plate were used. In the primary and secondary screens, 40 librarysiRNAs and 8 control siRNAs were transfected per plate, including four non-targeting control siRNAs (NTC), two siRNAs targeting cMet (MET) and twosiRNAs targeting MST1R. The confluent cell monolayer was wounded using a 96well stainless steel pin tool (V&P Scientific, Inc.), which introduced a singlehorizontal wound in each well. After wounding, cell debris was aspirated, cellswere washed once in migration medium (GIBCOH GlutaMAXTM supplementedwith 0.5% FBS), and medium was replaced with migration medium alone in orderto measure basal migration, or migration medium supplemented with 10 ng/mlHGF. Triplicate wells were used for both conditions. A picture of each well wasacquired using the semi-automated imaging system, Discovery-1. Cells wereplaced at 37 C, 10% CO2 atmosphere and allowed to migrate for 6 hours, afterwhich a second picture of the same area of the wound was taken. Representativegrey scale pictures of control transfected cells treated with 0.5% FBS alone or with0.5% FBS and HGF are depicted in supplementary material Fig. S1.

Calculation of wound healing speeds

Two grey-scale images were obtained per well using the Discovery-1 imagingsystem; one at time point zero and one at 6 hours. Resulting images were analysedusing a customised MetaMorph (Molecular Devices) journal, which allowed us toquantify the area surrounding the wound, generating two area values for each well.The rate of wound closure per hour was calculated based on the difference inwound size during the six hour migration time course. Mean values and s.e.m.swere calculated for basal and HGF-induced migration (supplementary materialFig. S1, column graphs, light and dark blue bars, respectively).

Calculation of HGF induction scores

Values from each of the three replicates for both basal and HGF-induced migrationwere averaged to give a single value per replicate. The original data from one halfplate (48 siRNAs) is displayed in supplementary material Fig. S2A. Plotting basalmigration speed values (x-axis) versus HGF-induced migration speed data (y-axis)on a dot plot graph reveals a strong correlation between the two data sets(supplementary material Fig. S2B). Using this correlation between these two datasets, for each observed basal motility value, a corresponding expected HGF-motility value was calculated. Most observed HGF-motility values are very closeto the calculated, expected values. However, a subset of hits affect the ligand-stimulated migration more or less strongly than they affect the basal migration,therefore decreasing or increasing the fold migration. These hits can bedistinguished from the rest of the values by appearing in the dot plot diagramsignificantly above or below the correlation line, as exemplified in supplementarymaterial Fig. S2B by the MET controls (red crosses), and MET present in thelibrary (circled). In contrast, the points representing the non-targeting negativecontrols (filled dark grey circles) and MST1R (red boxes) lie on the correlationline. To quantify this effect for each siRNA, an HGF induction score wascalculated using the formula depicted in supplementary material Fig. S2C,which takes into account the observed basal and HGF wound healing speeds aswell as the expected HGF wound healing speed. An induction score of 1 representsnormal HGF induction, values below 1 represent reduced HGF induction andvalues above 1 represent increased HGF induction. The bar diagram insupplementary material Fig. S2D displays the induction scores calculatedfrom the values depicted in supplementary material Fig. S2A. Again, the valuefor c-Met which was present on this exemplar subplate, is boxed. This analysis wascarried out separately for each transfected library subplate and finally, all inductionscores obtained during the primary screen were normalised to the average of allinduction scores obtained from cells transfected with non targeting control siRNA(NTC).

MTT assays

MTT [3-(4, 5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Sigma]powder was dissolved in PBS to a final concentration of 5 mg/ml. 10 ml was addedto each well of a 96-well plate, which contained the wounded cell monolayer in100 ml cell culture medium. Cells were incubated at 37 C, 10% CO2 for45 minutes. Medium was aspirated and formazan crystals were dissolved in 50 mlDMSO (Sigma). After 10 minutes incubation at 37 C, plates were placed on ashaker for an additional 10 minutes. Absorbance was read on a SpectraMax Plusplate reader (Molecular Devices), coupled to SoftMax Pro 4.7 software. The fourvalues obtained were averaged to give one MTT value per transfected siRNA.

Nuclei count

After carrying out the wound healing assay, transfected cells were fixed using3.7% formaldehyde. Fixed cells were stained with Hoechst 33342 and Alexa Fluor555-coupled phalloidin and washed three times with PBS. For each 96-well, threepictures were taken of the centre of the well including the wound edge usingCellomicsH ArrayScanH and subsequently analysed with ThermoFisher Scientific-designed BioApplications software. Obtained values for nuclei counts were

normalised to the values obtained for phalloidin that reflects the cell growth areawithin each picture. The obtained values were averaged to generate one nucleicount value per transfected siRNA.

Normalisation of primary data

Average values for basal motility, nuclei count and MTT obtained from eachtransfected subplate (40 library values and 8 control values) were normalised usingthe plate median (calculated from 40 library values). All values were then furthernormalised to the average value obtained from all cells transfected with nontargeting control siRNA. To obtain a viability score, the MTT scores and Nucleicount scores thus obtained were averaged. For each readout, thresholds weredefined based on the normal distribution of the values obtained for NTC-transfected cells, using the calculated average 62.576 (P50.005) standarddeviations as cut-off points.

Time-lapse video microscopy

Transfected cells growing on 96-well plates were wounded 60 hours posttransfection, and cell debris was washed once with migration medium, afterwhich migration medium alone, or migration medium supplemented with 10 ng/mlHGF was added. The images were obtained using the automated invert NikonEclipse TE2000-E microscope equipped with 106Nikon Plan Fluor objective andNikon iXonEM camera (AndorTM Technology). Plates were left overnight in anincubator box, which ensured equilibrated temperature and CO2 diffusion for thespecimen, and also reduced focus instability. One image per well was acquiredat 10-minute intervals, over a 12-hour period. Alternatively, transfected cellsgrowing on 24-well or 96-well Essen ImagelockTM plates were wounded using awound-making tool (Essen) 60 hours post transfection, cell debris was washed offand cells were incubated in migration medium alone or migration mediumcontaining 10 ng/ml HGF. Plates were incubated in the IncuCyteTM automatedimaging system and pictures were taken every 30 minutes over a 12-hour timeperiod.

Cell tracking

Images for each condition generated using time-lapse microscopy were stackedto create movies. Movies were produced in MetaMorph software (MolecularDevices) using Codec compression. Where pictures had been taken usingthe IncuCyteTM system, movies were generated using the IncuCyteTM software.Cell tracks were obtained manually using the CellTracker software (KineticImaging Ltd), which recorded the migration patterns of individual cells. At least 40cells were tracked per condition. Each experiment was performed on threeindependent occasions. The cell tracking measurements were incorporated into acustom made journal (created by Dr Daniel Zicha) in Mathematica 5.2 software(Wolfram Research), which provided speed measurement expressed in

micrometers per hour (mm/h). Motility values were normalised to those obtainedfrom control treated cells transfected with non-targeting control siRNA (NTC). Inthe case of ERK rescue experiments in HeLa-cells, only GFP-positive cells weretracked.

Transwell assays

Cell migration was also assessed using an 8 mm pore 24-well format Transwellchamber assay (Becton Dickinson Discovery Labware, Bedford, MA). Cellswere transfected using a reverse transfection protocol in 24-well plates. Seventy-two hours after the transfection, cells were harvested and resuspended ingrowth medium (GIBCOH GlutaMAXTM medium supplemented with 10% FBS).A 100 ml cell suspension was added to the top of each transwell chamber and cellswere allowed to migrate for 3 hours. Lower chambers contained growth mediumalone or growth medium supplemented with 10 ng/ml HGF. Each condition wascarried out in duplicates. Additionally 100 ml of cell suspensions were plated ontonormal 24-well plates, containing growth medium alone or growth mediumsupplemented with HGF. Following the migration time course, cells wereresuspended with EDTA and the number of cells in the lower chamber wasquantified with Cell Titer-GloH (Promega). Additionally, the number of cellsplated on the 24-well plates was quantified and the ratio of migrated cells wascalculated.

Calculation of HGF migration scores

For wound healing assays, tracking experiments and transwell assays carried outwhen only comparing a small number of siRNAs, the fold migration wascalculated by dividing the migration value obtained from HGF treated cells by the

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value obtained from control treated cells. The migration score for a given librarysiRNA was calculated as follows: (FOLDlib21)/(FOLDNTC21).

Immunoblotting

Transfected cells were harvested in 16 NuPAGE sample buffer (Invitrogen) 48or 72 hours post transfection and separated on NuPAGE 4–12% gradientpolyacrylamide gels according to manufacturer’s instructions. Separated proteinswere transferred onto a methanol-activated PVDF membrane and detectedusing the following primary antibodies: ERK1/2 (Santa Cruz Biotechnology), P-MET (Cell Signaling), P-ERK1/2 (Cell Signaling), MET (Santa CruzBiotechnology), Tubulin (Sigma), Paxillin (Santa Cruz), P-Paxillin (Abcam),P-S-GSK3 (Cell Signaling), P-Y-GSK3 (Santa Cruz). Detection was carried outemploying ECL, with visualisation using Hyperfilm (Amersham Biosciences) orwith the ImageQuantTM LAS 4000 mini system for quantitation (GE Healthcare).Densitometric analysis of immunoblots within the linear range was performedusing Image J 1.386software (Wayne Rasband, National Institutes of Health,Bethesda, MD). Background values were subtracted, and obtained values werenormalised to the tubulin loading control.

Monitoring of paxillin turnover

HeLa cells growing on glass bottom dishes were transfected with GFP-taggedpaxillin and two days later either incubated in 10% FCS or 10% FCS plus 20 ng/mlHGF. Live cell imaging was performed using total internal reflection fluorescence(TIRF) microscopy. Pictures were taken every minute over a 30-minute timeperiod. The resulting movies were used to determine the assembly and disassemblyrates of paxillin-containing focal adhesions as described in (Berginski et al., 2011),using the focal adhesion analysis server (http://faas.bme.unc.edu/). Data derivedfrom at least ten different movies were analysed. Only rates derived from curvefitswith R2.0.75 were included in the analysis.

Statistical analysis

Statistical significance was assessed by paired Student’s t-test, one-way Anova ortwo-way Anova followed by Bonferroni post hoc test. *P,0.05, **P,0.01,***P,0.001; ns, not significant.

AcknowledgementsWe thank the High-Throughput Screening Laboratory and the LightMicroscopy Laboratory at the LRI for help with the live-cell imagingand the Nikon Imaging Centre at the Institut Curie-CNRS for theirhelp with the TIRF microscopy experiments.

Author contributionsS.R. and M.D.R. designed the screen, S.R. wrote the manuscript, S.R.and M.M. carried out the primary screen and hit validation, C.R.carried out the TIRF experiments, S.L., C.R. and A.H. contributed tothe set up of the transwell assays and live cell imaging, S.K. and P.P.supervised the project and edited the manuscript.

FundingThis work was supported by Cancer Research UK; the DeutscheForschungsgemeinschaft (DFG) [grant number RA 1595/1-1]; andthe British Lung Foundation.

Supplementary material available online at

http://jcs.biologists.org/lookup/suppl/doi:10.1242/jcs.115832/-/DC1

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ERK2 mediates HGF-induced motility 2391

Fig. S1. Schematic representation of the set up of an siRNA-based high-throughput wound healing screen carried out in A549 NSCLC cells. Each library masterplate was divided into two half subplates; the scheme gives an overview of the transfection of one library subplate (shown in blue). A549 cells were transfected with the library siRNA and control siRNA pools and grown to confluency. For each siRNA pool, six wells on separate 96-well plates were transfected. Seventy two hours post transfection, wounding was carried out. Two sets of sample pictures taken of cells transfected with a non-targeting control siRNA are depicted. Pictures were taken directly and six hours after wounding on the Discovery-1, picture analysis was carried out using Metamorph software and the migration speed was calculated. Triplicate values were obtained from control treated and HGF-treated cells, and average wound healing speeds were calculated from the derived replicate values. Subsequently an MTT-assay was carried out on four of the six transfected samples and two samples were used for IF staining of the nuclei. Average nuclei counts and average MTT values were calculated.

0200004000060000

ctrl HGF

0.5% FCS

Day 4

Day1

average basal migration

speed

Nuclei count

0.5% FCS + HGF [10ng/ml]

average HGF migration

speed

Reverse transfection siRNAs

Wounding

MTT assay

picture analysis (metamorph)

picture acquisition (discovery)

Calculate migration speed =Δarea/Δtime

Figure S1

0h 6h

$"#!"#

0h 6h

0h $"#!"# $"# $"#0h 6h 0h 6h

Immunofluorescence Immunofluorescence

MTT Score

mig

ratio

n sp

eed

0 20000 40000 60000

0 0 0 0

mig

ratio

n sp

eed

p

0200004000060000

Fig. S2. Data analysis of the raw migration data. Data analysis carried out during the primary screen is displayed for one representative siRNA subplate. For each subplate, A549 cells were transfected with 48 siRNA pools, of which 8 were negative and positive control siRNAs. For each siRNA, six 96-wells on different plates were transfected and seventy-two hours post transfection, wound healing assays were carried out. Triplicate wound healing speeds were obtained for control treated and HGF-treated cells for each siRNA transfected and mean values and s.e.m.s were calculated. A: Wound healing speeds obtained from a subplate of library siRNAs transfected in the primary screen. Average migration speeds and s.e.m.s derived from triplicate values are displayed. Light blue bars represent values derived from control treated cells, dark blue bars represent values obtained from HGF-treated cells. The first 8 pairs of values represent the negative (non-targeting siRNA) and positive controls (MET, MST1R) included, followed by the 40 pairs of values from the transfected siRNA library. The displayed subset of siRNAs contains the siRNA pool targeting MET; the values obtained for this siRNA are boxed. B+C: Generation of the HGF-induction scores. To get a read out for siRNAs that specifically affect HGF-induced motility, for all transfected siRNAs, the obtained HGF migration speeds were blotted against the respective values obtained from control treated cells, revealing a strong correlation between both sets of values represented in the dot plot by the regression line (B). The R-value is displayed in the diagram. Light gray circles represent data from the library, values from NTC-transfected cells are displayed as dark gray circles, values for MST1R are displayed by red boxes, and red crosses represent values obtained for the positive control MET. Data points close to or on this line represent siRNAs, which do not specifically affect HGF-induced motility whereas data points above or below the regression line represent siRNAs that positively or negatively affect the HGF-response. The data point for MET which was included on this library subplate is circled. For each basal motility value obtained the respective value on the regression line can be calculated on basis of the correlation between both sets of data. For each pair of values an HGF-induction score was calculated by the formula displayed in the boxed area (C), taking into account the obtained (Vctrl, VHGF) as well as the calculated (Vcalc) motility values. A value of 1 means a given siRNA has no effect on HGF-inducibility whereas values below or above 1 point to a negative or positive effect on HGF-inducibility. D: HGF-induction scores for a subset of siRNAs transfected in the primary screen. From the raw migration data displayed in (A), induction scores were calculated as detailed in Material and Methods and described in (B). Induction scores are displayed in the same order as in (A). The box highlights the induction score calculated for the siRNA against MET.

0 20000 40000 60000 80000

100000

NTC

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TC

NTC

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TC

MET

M

ET

MST

1R

MST

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P3K

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AP4

K1

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K3

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P4K

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APK

13

MA

PK14

M

APK

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APK

4 M

APK

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APK

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APK

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APK

APK

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APK

APK

5 M

AR

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K

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MET

M

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1616

9 M

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4542

8 M

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4642

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4796

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5601

M

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8407

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MPP

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L1

MR

C2

MY

LK

MY

LK2

MY

O3A

M

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3B

NA

GK

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EK3

NEK

4 N

EK6

NEK

7 N

EK8

siRNA

ctrl HGF

0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75

NTC

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NTC

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MET

M

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MST

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1R

MA

P3K

9 M

AP4

K1

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P4K

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APK

13

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PK14

M

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APK

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APK

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M

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1616

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4542

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4642

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4796

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5601

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8407

M

PP1

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MPZ

L1

MR

C2

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LK

MY

LK2

MY

O3A

M

YO

3B

NA

GK

N

EK3

NEK

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EK6

NEK

7 N

EK8

siRNA

R2 = 0.78

0 10000 20000 30000 40000 50000 60000 70000 80000

0

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2000

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Figure S2

HGF induction score vHGF - vctrl vcalc - vctrl

D

C

A

B

library

NTC

MST1R

MET

wou

nd h

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wound healing speed (ctrl) [area units/hour]

HG

F in

duct

ion

scor

e w

ound

hea

ling

spee

d [(

HG

F)

[are

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Fig. S3. Effect of ERK2 knockdown by a single oligo on paxillin and MET phosphorylation. A+B: Strong correlation between knockdown efficiency of single ERK2-oligos and their effect on wound healing. The original ERK2 siRNA pool used in the primary screen was deconvoluted and single oligos were transfected into A549 cells. Total cellular lysates were generated 72 hours post transfection and the expression of ERK2 was monitored by Western Blot analysis. Representative blots are depicted in the left panel, blots were quantified as described in materials and methods and the relative ERK2-expression levels were calculated. Average values and s.e.m.s derived from three independent experiments are depicted. (A). Wound healing assays were carried out and HGF induction scores were generated as described in material and methods. Average values and s.e.m.s derived from three independent experiments are depicted (B). C: Knockdown of ERK2 with ERK2-oligo 1 or ERK2-oligo 4 prevents paxillin phosphorylation. A549 cells were transfected with a single siRNA oligo (E2-1, E2-4) or a non-targeting control (NTC). Cells were treated with HGF 48 hours post transfection for the indicated time or left untreated and total cellular lysates were generated. Lysates were analysed by Western blot as indicated. Representative blots are shown on the upper panel. Blots were quantified using image J software and values were normalised to values obtained for tubulin. Average normalised values and s.e.m.s from three independent experiments are depicted. To determine statistical significance 2-way anova was carried out, followed by Bonferroni post hoc test.

Figure S3

ERK2 Tubulin

O1 O2 O3 O4 NTC ERK2 siRNA

A

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on

***

*** ***

*** NTC O1 O2 O3 O4 B

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0 30 60 120 0 30 60 120 C NTC ERK2_4

0.00 0.25 0.50 0.75 1.00 1.25

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NTC ERK2-1 ERK2-4

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0.00 0.25 0.50 0.75 1.00 1.25

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NTC ERK2-1 ERK2-4

P-Pax

0.00 0.50 1.00 1.50 2.00 2.50 3.00

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NTC ERK2-1 ERK2-4

P-ERK1

0.00 0.25 0.50 0.75 1.00 1.25

0 30 60 120 0 30 60 120 0 30 60 120

NTC ERK2-1 ERK2-4

P-ERK2

0.00 0.25 0.50 0.75 1.00 1.25

0.00 0.25 0.50 0.75 1.00 1.25

0 30 60 120 0 30 60 120

Fig. S4. ERK2 knockdown prevents HGF-induced paxillin phosphorylation in various cell lines. CALU cells (A), SK-MES cells (B) or HeLa cells (C) were transfected with control siRNA (NTC) or siRNA targeting ERK2 or ERK1. 72 hours post transfection, cells were treated for 2 hours with HGF or left untreated, total cellular lysates were generated and analysed by Western blot as indicated. Representative blots are shown on the left panels. Blots were quantified using image J software and values were normalised to values obtained for tubulin. Average normalised values and s.e.m.s are depicted. To determine statistical significance, 2-way anova was carried out, followed by Bonferroni post hoc test.

Figure S4

ERK2 ERK1 NTC - + - + - +

CALU

HELA

SK-MES

- + - + - + NTC ERK1 ERK2

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P-ERK1 P-ERK2 P-PAX

P-ERK1 P-ERK2 P-PAX

P-ERK1 P-ERK2 P-PAX

0.00 0.50 1.00 1.50 2.00

NTC ERK1 ERK2 - + - + - +

NTC ERK1 ERK2 - + - + - +

0.00 0.25 0.50 0.75 1.00 1.25

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NTC ERK1 ERK2 - + - + - +

*** ***

***

0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50

NTC ERK1 ERK2 - + - + - +

0.00 0.25 0.50 0.75 1.00 1.25 1.50

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NTC ERK1 ERK2 NTC ERK1 ERK2 - + - + - + - + - + - +

ns **

NTC ERK1 ERK2 NTC ERK1 ERK2 NTC ERK1 ERK2 - + - + - + - + - + - + - + - + - + 0.00

0.50 1.00 1.50 2.00 2.50

0.00 0.25 0.50 0.75 1.00 1.25 1.50

*** *

**

0.00 0.25 0.50 0.75 1.00 1.25

rela

tive

sign

alre

lativ

e si

gnal

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sign

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Fig. S5. HGF regulates GSK3 activity. Cells were treated with HGF for the indicated time points and total cellular lysates were generated. Lysates were analysed by Western blot with the antibodies indicated. Representative blots are shown on the left panel. Blots were quantified using image J software and values were normalised to values obtained for tubulin. A: Average normalised values and s.e.m.s from five independent experiments are depicted. Statistical significance was determined using 1-way anova followed by Dunnett’s post hoc test. B: Prior to HGF-stimulation, cells were incubated for 20 min with LY294002 (LY) and then stimulated for the times indicated. Average normalised values and s.e.m.s from three independent experiments are depicted. Statistical significance was determined using the paired student’s t-test.

0 30 60 120 240 360

Tubulin

P-AKT

Figure S5

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P-AKT

0.00.20.40.60.81.01.2

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DMSO LY0 60 120 0 60120

ns*

P-Ser-GSK3 P-PAX

Fig. S6. ERK2 is more efficient than ERK1 in rescuing the effect of depletion of endogenous ERK2 on paxillin phosphorylation and single cell motility in HeLa cells. HeLa cells were transfected with control siRNA (NTC) or a single siRNA targeting ERK2 (E2-1). 24 hours post transfection, cells were transfected with cDNA encoding GFP, or HA-tagged ERK1 (E1HA) or ERK2 (E2HA). The next day, experiments were performed. A: ERK2 is more efficient than ERK1 in rescuing the effect of depletion of endogenous ERK2 on paxillin phosphorylation. Transfected cells were treated for 2 hours with HGF or left untreated, total cellular lysates were generated and analysed by Western blot as indicated. Representative blots are shown in the upper panels. Blots were quantified using image J software and values were normalised to values obtained for tubulin. To compare the efficiency of overexpressed ERK1 and ERK2, values obtained for fold P-paxillin were normalised using the values obtained for P-ERK upon HGF-stimulation. Average normalised values and s.e.m.s are depicted. To determine statistical significance, paired student’s t-test was carried out. B: ERK2 is more efficient than ERK1 in rescuing the effect of depletion of endogenous ERK2 on single cell motility. Transfected cells growing on 96-well plates were wounded and incubated in medium containing 0.5% FCS or 0.5FCS and HGF and video microscopy was performed. Single cell motility was determined by tracking single GFP-positive cells from 6 movies. Average single cell motility values and s.e.m.s are displayed. Statistical significance was determined by 1-way anova using the value obtained from HGF-treated NTC transfected cells as a reference.

Figure S6

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cDNA- + - + - + - +HGF

P-MET

P-PAXPAX

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Fold P-Paxctrl HGF

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GFP GFP E1HA E2HA NTC E2-1

P-MET ns

GFP GFP E1HA E2HA NTC E2-1

GFP GFP E1HA E2HA NTC E2-1

0.00 0.50 1.00 1.50 2.00 2.50

*

Fold P-Pax/P-ERK

Table S1 siRNA library HGF

induction score

basal motility

score

viability score

order number

gene locus

Accession number

AAK1 kinome 1.12 0.42 0.72 M-005300-00 22848 NM_014911

AATK kinome 0.72 0.87 1.03 M-005301-01 9625 XM_290778

ABL1 kinome 0.77 0.63 0.90 M-003100-01 25 NM_005157

ABL2 kinome 1.04 1.04 0.87 M-003101-01 27 NM_005158

ACK1 motility 0.68 0.70 0.84 M-003102-02 10188 NM_005781

ACK1 kinome 0.89 0.92 0.78 M-003102-02 10188 NM_005781

ACTN1 kinome 0.36 0.59 0.92 M-011195-00 87 NM_001102

ACTN2 kinome 0.78 1.05 0.92 M-011196-01 88 NM_001103

ACTN3 kinome 1.67 1.43 1.00 M-011197-01 89 NM_001104

ACTN4 kinome 1.41 1.09 0.86 M-011988-01 81 NM_004924

ACVR1 kinome 1.09 0.99 0.92 M-004924-01 90 NM_001105

ACVR1B kinome 0.89 0.87 1.00 M-004925-01 91 NM_004302

ACVR2 kinome 0.78 0.99 1.02 M-004926-01 92 NM_001616

ACVR2B kinome 1.07 0.76 0.97 M-004927-00 93 NM_001106

ACVRL1 kinome 0.84 1.44 1.00 M-005302-02 94 NM_000020

ADAM9 kinome 1.12 0.74 0.98 M-004504-00 8754 NM_003816

ADCK1 kinome 1.00 0.92 1.05 M-005303-02 57143 NM_020421

ADCK2 kinome 0.75 0.79 0.89 M-005304-01 90956 NM_052853

ADCK4 kinome 1.19 0.92 0.87 M-005305-01 79934 NM_024876

ADCK5 kinome 0.71 1.05 0.95 M-018919-00 203054 NM_174922

ADD1 kinome 1.14 0.61 1.02 M-009487-00 118 NM_001119

ADD2 kinome 0.88 0.85 1.00 M-009410-00 119 NM_001617

ADD3 kinome 0.70 0.78 0.92 M-008468-00 120 NM_016824

ADK kinome 1.24 0.75 0.77 M-004733-02 132 NM_001123

ADRA1A kinome 0.63 0.90 0.92 M-005419-00 148 NM_000680

ADRA1B kinome 0.75 0.60 0.92 M-005420-01 147 NM_000679

ADRB2 kinome 1.16 1.14 0.91 M-005426-01 154 NM_000024

ADRBK1 kinome 1.00 0.89 0.87 M-004325-01 156 NM_001619

ADRBK2 kinome 0.72 0.82 0.94 M-004326-00 157 NM_005160

ADRBK2 motility 0.84 0.82 0.99 M-004326-00 157 NM_005160

AGTR2 kinome 0.75 0.87 1.04 M-005429-01 186 NM_000686

AK1 kinome 1.31 0.55 0.89 M-006811-01 203 NM_000476

AK2 kinome 1.09 0.77 0.91 M-006812-00 204 NM_001625

AK3 kinome 0.77 1.38 1.00 M-006700-01 205 NM_013410

AK3L1 kinome 0.97 0.81 1.07 M-006701-00 50808 NM_016282

AK5 kinome 0.87 0.80 1.06 M-004897-01 26289 NM_012093

AK7 kinome 0.88 1.44 1.09 M-007257-00 122481 NM_152327

AKAP1 kinome 1.02 0.98 0.89 M-011426-00 8165 NM_003488

AKAP11 kinome 1.25 1.03 1.00 M-009277-00 11215 NM_016248

AKAP13 kinome 1.24 1.12 0.91 M-008868-00 11214 NM_006738

AKAP3 kinome 1.09 0.97 1.03 M-009765-00 10566 NM_006422

AKAP4 kinome 1.03 0.81 1.01 M-008442-00 8852 NM_003886

AKAP5 kinome 1.12 0.89 0.95 M-011954-00 9495 NM_004857

AKAP6 kinome 0.89 1.26 0.83 M-008497-00 9472 NM_004274

AKAP7 kinome 0.67 0.90 0.96 M-013371-00 9465 NM_004842

AKAP8 kinome 0.90 1.02 0.99 M-009656-00 10270 NM_005858

AKT1 kinome 0.62 0.62 0.91 M-003000-01 207 NM_005163

AKT2 kinome 0.84 0.81 0.85 M-003001-01 208 NM_001626

AKT3 kinome 0.89 0.69 1.08 M-003002-01 10000 NM_005465

ALK kinome 0.91 0.45 0.80 M-003103-02 238 NM_004304

ALK7 kinome 0.96 0.55 1.05 M-004929-01 130399 NM_145259

ALS2 motorpr 0.84 0.65 0.82 M-014168-01 57679 NM_020919

ALS2CR2 kinome 0.44 0.50 0.95 M-005306-00 55437 NM_018571

ALS2CR7 kinome 0.86 1.65 0.99 M-004685-01 65061 NM_139158

AMHR2 kinome 1.36 0.78 0.84 M-005307-00 269 NM_020547

ANGPT4 kinome 0.79 0.84 0.79 M-007803-00 51378 NM_015985

ANKK1 kinome 0.89 0.91 0.95 M-004930-00 255239 NM_178510

ANKRD3 kinome 0.80 1.12 0.99 M-005308-00 54101 NM_020639

APC2 motility 0.69 0.53 0.88 M-009847-00 10297 NM_005883

APEG1 kinome 0.52 0.86 0.94 M-007687-00 10290 NM_005876

APG10L kinome 0.78 0.82 0.92 M-019426-00 83734 NM_031482

APG12L kinome 0.96 0.90 1.02 M-010212-00 9140 NM_004707

APG3 kinome 0.59 1.06 0.95 M-015375-00 64422 NM_022488

APG5L kinome 0.76 0.99 0.96 M-004374-01 9474 NM_004849

APG7L kinome 0.58 1.04 1.01 M-020112-00 10533 NM_006395

APPL kinome 1.38 0.66 0.83 M-005138-00 26060 NM_012096

ARAF1 kinome 1.11 1.03 0.99 M-003563-02 369 NM_001654

ARHE motility 0.69 0.66 0.94 M-007794-01 390 NM_005168

ARHE motility 0.73 0.68 0.94 M-007794-01 390 NM_005168

ARHGDIB motility 0.88 1.02 1.08 M-010893-01 397 NM_001175

ARHN motility 1.03 1.30 0.97 M-009727-00 8153 NM_005440

ARK5 kinome 1.03 1.16 1.08 M-004931-00 9891 NM_014840

ASK kinome 0.68 0.71 1.09 M-004165-01 10926 NM_006716

ASP kinome 0.85 1.18 1.03 M-013023-00 83853 NM_031916

ATM kinome 0.79 1.11 1.19 M-003201-01 472 NM_000051

ATR kinome 1.24 0.85 0.85 M-003202-04 545 NM_001184

AURKB kinome 1.54 0.86 0.87 M-003326-02 9212 NM_004217

AURKC kinome 1.00 0.46 0.88 M-019573-01 6795 NM_003160

AVPR1A kinome 1.94 1.05 0.92 M-003631-01 552 NM_000706

AVPR1B kinome 1.45 0.59 0.78 M-005431-00 553 NM_000707

AXL kinome 1.01 0.85 0.90 M-003104-01 558 NM_001699

AZU1 kinome 0.36 0.52 0.75 M-008914-00 566 NM_001700

BAK1 kinome 0.44 0.58 0.93 M-003305-01 578 NM_001188

BAX kinome 0.92 0.58 1.11 M-003308-00 581 NM_004324

BCKDK kinome 0.78 0.91 0.79 M-004932-00 10295 NM_005881

BCL2 motility 0.77 0.46 0.71 M-003307-04 596 NM_000633

BCL2 motility 1.20 0.39 0.74 M-003307-04 596 NM_000633

BCL2 kinome 1.22 0.81 0.80 M-003307-04 596 NM_000633

BCL2A1 kinome 0.77 1.09 1.06 M-003306-00 597 NM_004049

BCL2L1 kinome 1.07 0.45 0.93 M-003458-00 598 NM_138578

BCL2L2 kinome 1.15 1.02 0.82 M-004384-01 599 NM_004050

BCR kinome 0.74 0.79 0.76 M-003875-04 613 NM_004327

BDKRB2 kinome 0.72 0.94 0.93 M-005436-00 624 NM_000623

BECN1 kinome 0.51 0.66 0.90 M-010552-00 8678 NM_003766

BICD1 motorpr 0.96 1.08 0.83 M-019496-01 636 NM_001714

BICD2 motorpr 0.70 0.75 0.97 M-014060-01 23299 NM_015250

BIRC2 kinome 0.97 0.88 0.97 M-004390-01 329 NM_001166

BIRC3 kinome 1.29 0.84 0.96 M-004099-02 330 NM_001165

BIRC4 kinome 0.90 0.49 0.91 M-004098-00 331 NM_001167

BIRC7 kinome 1.04 1.06 0.96 M-004391-01 79444 NM_022161

BLK kinome 0.91 0.38 0.83 M-003105-02 640 NM_001715

BLNK kinome 0.70 0.70 0.98 M-020353-00 29760 NM_013314

BMP2K kinome 0.71 0.96 0.93 M-005071-00 55589 NM_017593

BMPR1A kinome 0.69 0.88 0.93 M-004933-03 657 NM_004329

BMPR1B kinome 1.19 0.68 0.81 M-004934-00 658 NM_001203

BMPR2 kinome 0.94 0.41 0.91 M-005309-01 659 NM_001204

BMX motility 1.09 1.36 1.00 M-003106-02 660 NM_001721

BMX kinome 1.17 1.10 0.86 M-003106-01 660 NM_001721

BRAF kinome 1.52 0.69 0.52 M-003460-01 673 NM_004333

BRAF motility 1.71 0.66 0.64 M-003460-01 673 NM_004333

BRD2 kinome 0.75 0.90 0.87 M-004935-00 6046 NM_005104

BRDT kinome 0.77 1.17 0.91 M-004938-00 676 NM_001726

BTK kinome 0.85 0.95 0.98 M-003107-01 695 NM_000061

BUB1 kinome 0.51 0.86 0.98 M-004102-00 699 NM_004336

BUB1B kinome 0.55 0.89 0.91 M-004101-00 701 NM_001211

C14ORF20 kinome 0.79 0.83 0.95 M-005310-00 283629 NM_174944

C20ORF23 motorpr 0.71 0.57 0.77 M-009495-01 55614 NM_024704

C20ORF64 kinome 0.73 1.04 0.96 M-003108-01 112858 NM_033550

C20ORF97 kinome 0.49 1.05 0.87 M-003754-01 57761 NM_021158

C6ORF102 motorpr 0.65 0.86 0.94 M-016448-00 221458 NM_145027

C6ORF199 kinome 0.86 0.59 0.89 M-007254-00 221264 NM_145025

C7ORF16 kinome 0.83 1.06 0.82 M-018324-00 10842 NM_006658

C8FW kinome 0.82 1.16 0.86 M-003633-00 10221 NM_025195

C9ORF12 kinome 0.69 0.57 0.81 M-006703-01 64768 NM_022755

C9ORF48 motorpr 1.32 0.68 0.81 M-010459-00 347240 NM_194313

CALM3 kinome 0.68 0.80 0.93 M-019939-00 808 NM_005184

CAMK1 kinome 0.85 0.73 0.85 M-004940-00 8536 NM_003656

CAMK1D kinome 0.76 0.81 0.95 M-004946-00 57118 NM_020397

CAMK1G kinome 0.71 1.10 0.96 M-004941-00 57172 NM_020439

CAMK2A kinome 0.75 0.93 0.98 M-004942-00 815 NM_015981

CAMK2B kinome 0.96 1.20 1.06 M-004943-01 816 NM_001220

CAMK2D kinome 1.32 0.56 0.96 M-004042-02 817 NM_001221

CAMK2G kinome 0.96 1.02 0.99 M-004536-01 818 NM_001222

CAMK4 kinome 0.93 1.06 0.92 M-004944-00 814 NM_001744

CAMKK1 kinome 1.00 1.57 0.96 M-004912-00 84254 NM_172206

CAMKK2 kinome 1.17 1.07 0.92 M-004842-00 10645 NM_006549

CARD10 kinome 1.15 0.92 1.00 M-004395-00 29775 NM_014550

CARD14 kinome 0.75 0.90 0.96 M-004397-00 79092 NM_024110

CARK kinome 0.91 1.04 0.93 M-005013-00 51086 NM_015978

CARKL kinome 0.51 1.26 0.93 M-006815-00 23729 NM_013276

CASK kinome 0.15 0.18 0.74 M-005311-00 8573 NM_003688

CASK motility 0.79 0.32 0.85 M-005311-01 8573 NM_003688

CCL2 kinome 0.76 1.38 0.82 M-007831-00 6347 NM_002982

CCL4 kinome 0.76 0.66 0.98 M-007843-00 6351 NM_002984

CCRK kinome 0.20 0.28 0.66 M-004686-01 23552 NM_012119

CD3E kinome 0.86 0.69 0.79 M-003775-01 916 NM_000733

CD4 kinome 1.32 0.86 0.90 M-005234-00 920 NM_000616

CD7 kinome 1.24 0.84 0.90 M-013630-00 924 NM_006137

CDACD1 kinome 1.02 0.91 0.90 M-007732-00 81602 NM_030911

CDC2 motility 0.08 1.08 0.44 M-003224-03 983 NM_001786

CDC2 kinome 0.49 0.86 0.53 M-003224-03 983 NM_001786

CDC2 motility 0.91 0.75 0.48 M-003224-03 983 NM_001786

CDC2L1 motility 0.49 0.49 0.80 M-004687-01 984 NM_001787

CDC2L1 motility 0.70 0.58 0.77 M-004687-01 984 NM_001787

CDC2L1 kinome 0.87 1.02 1.00 M-004687-01 984 NM_001787

CDC2L2 kinome 0.64 0.72 0.83 M-004026-01 985 NM_024011

CDC2L5 kinome 1.08 1.36 0.89 M-004688-00 8621 NM_003718

CDC42 motility 0.56 0.89 1.03 M-005057-00 998 NM_001791

CDC42 motility 0.72 0.64 0.73 M-005057-00 998 NM_001791

CDC42 kinome 0.84 0.89 0.86 M-005057-00 998 NM_001791

CDC42BPA kinome 0.91 1.14 0.95 M-003814-02 8476 NM_003607

CDC42BPB kinome 0.96 1.40 1.01 M-004075-00 9578 NM_006035

CDC7L1 kinome 0.89 0.89 0.93 M-003234-01 8317 NM_003503

CDK10 kinome 0.91 1.00 0.91 M-003235-03 8558 NM_003674

CDK11 kinome 1.01 0.55 0.83 M-004689-00 23097 NM_015076

CDK2 kinome 0.92 1.04 0.98 M-003236-04 1017 NM_001798

CDK3 kinome 0.68 1.09 1.00 M-003237-01 1018 NM_001258

CDK4 kinome 1.13 0.70 0.89 M-003238-02 1019 NM_000075

CDK5 kinome 0.86 0.99 1.04 M-003239-01 1020 NM_004935

CDK5R1 kinome 1.34 0.46 0.90 M-008988-00 8851 NM_003885

CDK5R2 kinome 0.55 0.41 0.94 M-008885-00 8941 NM_003936

CDK5RAP1 kinome 1.58 1.01 0.88 M-013297-00 51654 NM_016082

CDK5RAP3 kinome 1.10 0.81 0.98 M-012957-00 80279 NM_025197

CDK6 kinome 1.02 0.55 0.87 M-003240-02 1021 NM_001259

CDK7 kinome 1.23 1.10 0.98 M-003241-01 1022 NM_001799

CDK8 kinome 1.29 1.48 0.88 M-003242-02 1024 NM_001260

CDK9 kinome 0.53 0.58 0.83 M-003243-02 1025 NM_001261

CDKL1 kinome 1.03 0.77 0.94 M-004323-01 8814 NM_004196

CDKL2 kinome 0.64 1.16 0.91 M-004797-00 8999 NM_003948

CDKL3 kinome 0.78 0.93 1.14 M-004798-00 51265 NM_016508

CDKL5 kinome 0.64 1.30 0.87 M-004799-02 6792 NM_003159

CDKN1A kinome 0.89 0.90 1.16 M-003471-00 1026 NM_000389

CDKN1B kinome 1.04 0.65 0.93 M-003472-00 1027 NM_004064

CDKN1C kinome 1.08 0.76 0.82 M-003244-03 1028 NM_000076

CDKN2A kinome 0.77 0.71 0.97 M-011007-00 1029 NM_000077

CDKN2B kinome 0.75 1.06 0.98 M-003245-01 1030 NM_004936

CDKN2C kinome 0.90 1.19 1.07 M-003246-01 1031 NM_001262

CDKN2D kinome 1.10 1.18 0.97 M-003247-02 1032 NM_001800

CDKN3 kinome 1.30 0.62 0.77 M-003879-00 1033 NM_005192

CENPE motorpr 0.73 0.15 0.50 M-003252-01 1062 NM_001813

CERK kinome 0.94 0.99 0.93 M-004061-00 64781 NM_022766

CFL1 motility 1.21 0.70 0.76 M-012707-00 1072 NM_005507

CFL1 motility 1.27 0.57 0.75 M-012707-00 1072 NM_005507

CFLAR kinome 0.79 1.01 0.88 M-003772-05 8837 NM_003879

CGEF2 kinome 1.20 0.50 1.03 M-009511-00 11069 NM_007023

CHEK1 kinome 0.96 0.81 0.88 M-003255-02 1111 NM_001274

CHEK2 kinome 1.23 0.88 0.99 M-003256-05 11200 NM_007194

CHK kinome 1.12 0.62 0.77 M-006704-00 1119 NM_001277

CHKL kinome 1.08 0.75 0.99 M-006705-00 1120 NM_005198

CHRM1 kinome 1.18 0.79 0.93 M-005462-00 1128 NM_000738

CHUK motility 0.73 0.68 1.04 M-003473-01 1147 NM_001278

CHUK kinome 0.93 0.69 0.90 M-003473-01 1147 NM_001278

CINP kinome 0.54 0.75 1.01 M-013591-00 51550 NM_032630

CIT kinome 0.96 0.82 0.90 M-004613-00 11113 NM_007174

CKB kinome 0.75 1.25 0.99 M-006706-01 1152 NM_001823

CKM kinome 1.04 0.40 1.00 M-006707-00 1158 NM_001824

CKMT1 kinome 1.09 0.59 0.74 M-006708-00 1159 NM_020990

CKMT2 kinome 0.93 0.91 0.86 M-006709-00 1160 NM_001825

CKS1B kinome 0.97 1.08 0.97 M-004586-00 1163 NM_001826

CKS2 kinome 0.97 0.78 0.94 M-007678-00 1164 NM_001827

CLK1 kinome 1.14 1.31 1.12 M-004800-00 1195 NM_004071

CLK2 kinome 1.08 0.72 1.23 M-004801-01 1196 NM_001291

CLK3 kinome 0.97 0.75 0.95 M-004802-01 1198 NM_001292

CLK4 kinome 0.78 1.20 0.91 M-004803-00 57396 NM_020666

CNK kinome 0.55 0.50 0.82 M-003257-02 1263 NM_004073

CNK1 kinome 0.89 0.49 0.55 M-012217-00 10256 NM_006314

COL4A3BP kinome 1.54 0.80 1.09 M-012101-00 10087 NM_005713

COPB2 kinome -0.05 0.12 0.38 M-019847-00 9276 NM_004766

CRK7 kinome 1.06 0.88 0.88 M-004031-02 51755 NM_016507

CRKL kinome 1.08 0.81 1.01 M-012023-01 1399 NM_005207

CSF1R kinome 1.02 0.67 0.77 M-003109-01 1436 NM_005211

CSK kinome 0.96 0.76 0.82 M-003110-01 1445 NM_004383

CSNK1A1 kinome 0.80 0.70 0.97 M-003957-03 1452 NM_001892

CSNK1D kinome 0.42 0.71 0.82 M-003478-00 1453 NM_001893

CSNK1E kinome 0.77 0.97 0.86 M-003479-01 1454 NM_001894

CSNK1G1 kinome 1.17 0.57 0.92 M-004666-01 53944 NM_022048

CSNK1G2 kinome 1.00 1.09 0.96 M-004678-00 1455 NM_001319

CSNK1G3 kinome 0.76 0.94 0.95 M-004679-01 1456 NM_004384

CSNK2A1 kinome 0.91 1.07 0.94 M-003475-00 1457 NM_001895

CSNK2A2 kinome 1.12 0.83 0.88 M-004752-00 1459 NM_001896

CSNK2B kinome 0.98 1.61 1.01 M-007679-00 1460 NM_001320

CTTN motility 0.67 1.04 0.89 M-010508-00 2017 NM_005231

CTTN motility 0.93 1.00 0.98 M-010508-00 2017 NM_005231

CUTL1 kinome 1.08 1.16 0.89 M-011635-00 1523 NM_001913

CXCL10 kinome 1.11 0.82 0.93 M-007871-00 3627 NM_001565

D2LIC motorpr 1.05 0.82 0.75 M-020872-00 51626 NM_015522

DAPK1 kinome 1.02 0.48 0.73 M-004417-02 1612 NM_004938

DAPK2 kinome 0.73 0.86 0.89 M-004418-02 23604 NM_014326

DAPK3 kinome 0.89 0.99 0.96 M-004947-00 1613 NM_001348

DCAMKL1 kinome 0.90 0.89 0.91 M-004884-01 9201 NM_004734

DCK kinome 0.86 1.12 1.01 M-006710-00 1633 NM_000788

DCTN1 motorpr 1.12 0.66 0.93 M-012874-00 1639 NM_004082

DCTN2 motorpr 1.16 1.10 1.07 M-012218-00 10540 NM_006400

DCTN3 motorpr 1.02 0.72 1.08 M-012365-00 11258 NM_007234

DCTN4 motorpr 1.10 0.69 1.05 M-015395-00 51164 NM_016221

DCTN6 motorpr 1.14 0.84 0.80 M-012302-00 10671 NM_006571

DDR1 motility 1.09 0.49 0.79 M-003111-03 780 NM_001954

DDR1 kinome 1.11 0.72 0.86 M-003111-02 780 NM_001954

DDR2 kinome 1.31 1.02 0.84 M-003112-02 4921 NM_006182

DGKA kinome 1.02 0.72 0.84 M-006711-01 1606 NM_001345

DGKB kinome 1.16 0.83 0.93 M-006712-01 1607 NM_004080

DGKD kinome 0.94 1.16 0.94 M-006713-02 8527 NM_003648

DGKE kinome 1.33 0.85 0.88 M-011493-00 8526 NM_003647

DGKG kinome 0.97 1.10 0.93 M-006715-00 1608 NM_001346

DGKI kinome 0.89 1.01 0.92 M-006717-00 9162 NM_004717

DGKQ kinome 0.72 0.74 1.02 M-005079-00 1609 NM_001347

DGKZ kinome 0.72 1.53 0.97 M-006718-00 8525 NM_003646

DGUOK kinome 0.96 0.75 0.90 M-006719-02 1716 NM_001929

DIAPH1 motility 1.05 1.08 1.03 M-010347-01 1729 NM_005219

DIAPH2 motility 0.97 0.78 1.02 M-012029-00 1730 NM_006729

DIAPH3 motility 0.66 1.04 1.02 M-018997-00 81624 NM_030932

DIAPH3 motility 1.03 1.12 1.22 M-018997-00 81624 NM_030932

DKFZP434C131

kinome 1.04 0.93 1.04 M-004949-00 25989 XM_044630

DKFZP434C1418

kinome 1.09 0.70 0.86 M-005313-01 285220 NM_173655

DKFZP586B1621

kinome 1.02 0.87 1.00 M-006808-00 26007 NM_015533

DKFZP761P0423

kinome 0.93 1.09 1.06 M-025870-00 157285 XM_291277

DKFZp761P1010

kinome 0.86 0.87 0.97 M-003113-01 55359 NM_018423

DLC1 motility 0.45 0.91 1.02 M-008713-00 10395 NM_006094

DLC2 motorpr 1.65 0.39 0.65 M-006493-01 140735 NM_080677

DLG1 kinome 0.97 0.97 0.97 M-009415-00 1739 NM_004087

DLG2 kinome 0.85 1.17 1.04 M-011252-00 1740 NM_001364

DLG3 kinome 1.06 0.95 0.98 M-009462-00 1741 NM_021120

DLG4 kinome 0.97 1.13 1.01 M-007882-00 1742 NM_001365

DMD kinome 1.01 1.08 0.97 M-011809-00 1756 NM_000109

DMPK kinome 1.05 0.92 0.85 M-004637-00 1760 NM_004409

DNAJC3 kinome 0.90 0.79 0.94 M-012251-00 5611 NM_006260

DNCI1 motorpr 1.72 0.79 0.93 M-019799-00 1780 NM_004411

DNCI2 motorpr 1.06 0.27 0.64 M-012574-00 1781 NM_001378

DNCL1 motorpr 0.52 0.73 0.67 M-005281-01 8655 NM_003746

DNCL2A motorpr 1.42 1.08 1.04 M-010586-01 83658 NM_014183

DNCL2B motorpr 0.73 0.93 1.04 M-015304-00 83657 NM_130897

DNCLI1 motorpr 0.99 1.05 0.99 M-021045-00 51143 NM_016141

DNCLI2 motorpr 0.69 1.07 0.90 M-020110-00 1783 NM_006141

DOK1 kinome 0.92 0.96 1.00 M-011254-00 1796 NM_001381

DTYMK kinome 0.98 1.42 1.05 M-006720-00 1841 NM_012145

DTYMK motility 1.26 1.17 1.14 M-006720-00 1841 NM_012145

DUSP1 kinome 0.87 1.03 0.96 M-003484-02 1843 NM_004417

DUSP10 kinome 0.74 1.00 0.99 M-003965-01 11221 NM_144728

DUSP2 kinome 0.94 0.58 0.65 M-003565-00 1844 NM_004418

DUSP22 kinome 0.66 1.06 0.96 M-004517-00 56940 NM_020185

DUSP4 kinome 1.20 0.93 1.01 M-003963-01 1846 NM_057158

DUSP5 kinome 1.62 0.52 0.77 M-003566-00 1847 NM_004419

DUSP6 kinome 0.87 0.81 1.09 M-003964-00 1848 NM_001946

DUSP7 kinome 0.58 0.61 1.00 M-003567-00 1849 NM_001947

DUSP8 kinome 0.84 1.00 0.89 M-003568-00 1850 NM_004420

DYNC1H1 motorpr 0.69 1.02 0.85 M-006828-00 1778 NM_001376

DYRK1A kinome 0.79 0.92 1.07 M-004805-00 1859 NM_001396

DYRK1B kinome 1.01 0.76 0.99 M-004806-01 9149 NM_004714

DYRK2 kinome 1.08 1.14 1.02 M-004730-02 8445 NM_003583

DYRK3 kinome 0.58 1.37 1.00 M-004731-00 8444 NM_003582

DYRK4 kinome 1.05 0.71 0.98 M-004732-01 8798 NM_003845

EDN2 kinome 0.54 0.89 1.04 M-017723-00 1907 NM_001956

EEF2K kinome 0.68 0.54 0.83 M-004950-00 29904 NM_013302

EGFR kinome 1.42 0.88 0.90 M-003114-01 1956 NM_005228

EIF2AK3 kinome 0.94 0.81 0.97 M-004883-01 9451 NM_004836

EIF2AK4 kinome 0.93 0.80 0.97 M-005314-00 27104 XM_031612

EKI1 kinome 0.87 0.49 0.81 M-006721-00 55500 NM_018638

EPAC kinome 0.88 0.76 0.80 M-007676-00 10411 NM_006105

EPHA1 kinome 0.64 0.94 0.84 M-003115-02 2041 NM_005232

EPHA2 kinome 1.28 0.71 0.84 M-003116-01 1969 NM_004431

EPHA3 kinome 1.10 0.52 0.78 M-003117-01 2042 NM_005233

EPHA4 kinome 0.74 0.66 0.90 M-003118-01 2043 NM_004438

EPHA4 motility 0.94 0.67 0.94 M-003118-01 2043 NM_004438

EPHA5 kinome 0.85 0.88 0.88 M-005315-01 2044 NM_004439

EPHA7 kinome 0.80 1.03 0.84 M-003119-01 2045 NM_004440

EPHA8 kinome 1.01 1.01 0.93 M-003120-02 2046 NM_020526

EPHB1 kinome 0.55 0.66 0.99 M-003121-01 2047 NM_004441

EPHB2 kinome 0.87 0.77 0.91 M-003122-01 2048 NM_004442

EPHB3 kinome 1.09 0.86 0.90 M-003123-01 2049 NM_004443

EPHB4 kinome 0.68 1.09 1.03 M-003124-01 2050 NM_004444

EPHB6 kinome 1.12 0.98 0.77 M-003125-01 2051 NM_004445

ERBB2 kinome 0.77 0.90 0.94 M-003126-01 2064 NM_004448

ERBB3 kinome 1.17 0.91 0.84 M-003127-01 2065 NM_001982

ERBB4 kinome 0.93 1.01 0.79 M-003128-02 2066 NM_005235

ERK8 kinome 0.74 0.90 0.97 M-004807-00 225689 NM_139021

ERN1 kinome 0.96 0.90 0.82 M-004951-00 2081 NM_001433

EVI1 kinome 0.88 0.85 0.82 M-006530-00 2122 NM_005241

FADD motility 0.93 1.23 1.26 M-003800-01 8772 NM_003824

FADD motility 1.01 1.19 1.20 M-003800-01 8772 NM_003824

FASTK kinome 0.85 0.73 0.87 M-005317-02 10922 NM_006712

FER kinome 0.81 0.79 0.88 M-003129-01 2241 NM_005246

FES kinome 0.59 1.18 0.89 M-003130-01 2242 NM_002005

FGFR1 kinome 0.96 0.84 0.91 M-003131-01 2260 NM_000604

FGFR2 motility 0.78 0.83 0.74 M-003132-01 2263 NM_000141

FGFR2 kinome 0.92 0.88 0.83 M-003132-01 2263 NM_000141

FGFR2 motility 1.19 1.20 0.99 M-003132-01 2263 NM_000141

FGFR3 kinome 1.02 0.66 0.72 M-003133-01 2261 NM_000142

FGFR4 kinome 1.14 0.83 0.83 M-003134-01 2264 NM_002011

FGR kinome 1.33 0.56 0.67 M-003135-02 2268 NM_005248

FLJ10074 kinome 0.88 1.04 0.98 M-005318-00 55681 NM_017988

FLJ10157 motorpr 0.59 0.33 0.54 M-022129-00 55083 XM_371354

FLJ10761 kinome 0.73 0.86 0.89 M-005078-00 55224 NM_018208

FLJ10842 kinome 1.03 0.93 0.97 M-007256-00 55750 NM_018238

FLJ11149 kinome 1.17 0.93 0.96 M-007260-00 55312 NM_018339

FLJ12476 kinome 0.94 0.83 0.94 M-007726-00 64799 NM_022784

FLJ13052 kinome 0.65 0.82 0.84 M-006318-00 65220 NM_023018

FLJ14813 kinome 0.98 1.07 1.00 M-004020-01 84930 NM_032844

FLJ20574 kinome 0.88 0.67 0.86 M-005319-00 54986 NM_017886

FLJ22002 kinome 0.98 0.83 0.99 M-007253-00 79896 NM_024838

FLJ23074 kinome 1.02 1.13 0.92 M-004843-00 80122 NM_025052

FLJ23119 kinome 0.80 0.81 0.93 M-005320-01 79705 NM_024652

FLJ23356 kinome 0.77 0.90 0.94 M-005321-00 84197 NM_032237

FLJ25006 kinome 0.52 1.08 0.95 M-004624-00 124923 NM_144610

FLJ32685 kinome 1.05 0.84 0.91 M-004052-02 152110 NM_152534

FLJ32704 kinome 0.89 0.92 0.98 M-006803-00 158067 NM_152572

FLJ32752 motorpr 1.56 1.06 1.12 M-015858-00 144132 NM_144666

FLJ34389 kinome 1.19 0.94 0.93 M-005326-00 197259 NM_152649

FLJ35107 kinome 1.08 0.67 1.03 M-007727-00 348825 NM_182629

FLJ37300 motorpr 0.66 0.68 0.97 M-004966-00 124602 NM_153209

FLJ40427 motorpr 0.42 0.73 0.82 M-017812-00 201625 NM_178504

FLJ46675 motorpr 0.82 1.60 1.05 M-030838-00 146754 XM_085578

FLNA kinome 0.34 1.38 1.03 M-012579-00 2316 NM_001456

FLNB kinome 1.19 1.26 1.03 M-020175-00 2317 NM_001457

FLT1 kinome 0.96 0.79 0.97 M-003136-01 2321 NM_002019

FLT3 kinome 1.00 0.86 0.82 M-003137-01 2322 NM_004119

FLT4 motility 0.99 0.53 0.84 M-003138-01 2324 NM_002020

FLT4 kinome 1.08 0.61 0.84 M-003138-01 2324 NM_002020

FLT4 motility 1.22 0.47 0.86 M-003138-01 2324 NM_002020

FMNL1 motility 0.64 0.78 0.89 M-019176-00 752 NM_005892

FMNL2 motility 1.02 1.05 1.16 M-031993-00 114793 NM_052905

FMNL3 motility 0.82 0.47 0.75 M-019007-00 91010 NM_175736

FMNL3 motility 1.15 0.73 1.08 M-019007-00 91010 NM_175736

FN3K kinome 0.70 1.06 0.95 M-006724-00 64122 NM_022158

FN3KRP kinome 0.67 0.61 0.88 M-006817-00 79672 NM_024619

FRAP1 kinome 0.53 0.42 0.75 M-003008-01 2475 NM_004958

FRDA kinome 0.82 0.97 1.06 M-006691-00 2395 NM_000144

FRK kinome 0.89 1.07 0.95 M-003139-01 2444 NM_002031

FUK kinome 1.01 0.70 0.73 M-007255-00 197258 NM_145059

FYB kinome 1.09 0.99 1.03 M-020174-00 2533 NM_001465

FYN kinome 0.89 0.88 1.03 M-003140-03 2534 NM_002037

GABARAP kinome 1.10 0.44 0.84 M-012368-00 11337 NM_007278

GABARAPL2

kinome 0.88 0.89 0.88 M-006853-00 11345 NM_007285

GAK kinome 0.89 0.79 0.86 M-005005-00 2580 NM_005255

GALK1 kinome 0.85 0.74 0.97 M-007728-00 2584 NM_000154

GALK2 kinome 0.76 0.92 0.79 M-006725-00 2585 NM_002044

GAP43 kinome 0.92 0.99 0.99 M-011663-00 2596 NM_002045

GCK kinome 0.77 0.81 1.02 M-010819-00 2645 NM_000162

GFRA2 kinome 1.17 0.88 0.92 M-007914-00 2675 NM_001495

GK kinome 0.72 0.98 1.01 M-006727-01 2710 NM_000167

GK2 kinome 0.81 0.97 0.99 M-015091-01 2712 NM_033214

GMFB kinome 1.09 1.06 1.09 M-019100-00 2764 NM_004124

GMFG kinome 1.13 0.76 1.03 M-019878-00 9535 NM_004877

GNE kinome 1.05 0.99 1.03 M-006729-00 10020 NM_005476

GPRK2L kinome 0.78 0.99 0.86 M-004625-00 2868 NM_005307

GPRK5 kinome 1.03 1.03 0.82 M-004626-00 2869 NM_005308

GPRK6 kinome 0.85 0.75 0.91 M-004627-01 2870 NM_002082

GPRK7 kinome 1.09 0.69 0.95 M-004628-00 131890 NM_139209

GRB2 motility 0.99 0.61 0.76 M-019220-00 2885 NM_002086

GSG2 kinome 1.05 0.98 0.96 M-005327-00 83903 NM_031965

GSK3A motility 1.10 0.98 0.94 M-003009-01 2931 NM_019884

GSK3A motility 1.13 1.01 0.91 M-003009-01 2931 NM_019884

GSK3A kinome 1.16 0.85 0.89 M-003009-01 2931 NM_019884

GSK3B kinome 0.82 0.95 0.96 M-003010-03 2932 NM_002093

GTF2H1 kinome 0.91 0.81 1.04 M-010924-00 2965 NM_005316

GUCY2C kinome 0.74 0.93 1.03 M-005328-00 2984 NM_004963

GUCY2D kinome 0.89 0.70 0.85 M-005329-00 3000 NM_000180

GUCY2F kinome 1.03 0.97 1.03 M-004515-01 2986 NM_001522

GUK1 kinome 0.61 0.94 0.96 M-006734-00 2987 NM_000858

H11 kinome 1.41 0.73 0.98 M-005006-00 26353 NM_014365

HAK kinome 1.07 0.89 1.01 M-005330-01 115701 NM_052947

HCK kinome 0.65 0.71 0.94 M-003141-01 3055 NM_002110

HIPK1 kinome 1.13 1.01 1.00 M-004809-02 204851 NM_152696

HIPK2 kinome 0.86 0.88 0.86 M-003266-03 28996 NM_022740

HIPK3 kinome 0.66 0.32 0.93 M-004810-00 10114 NM_005734

HIPK4 kinome 0.87 0.95 1.05 M-004808-02 147746 NM_144685

HK1 kinome 0.70 0.59 0.87 M-006820-01 3098 NM_000188

HK2 kinome 0.95 0.52 0.96 M-006735-01 3099 NM_000189

HK3 kinome 0.94 0.89 0.97 M-006736-00 3101 NM_002115

HRAS kinome 0.98 0.66 0.72 M-004142-00 3265 NM_005343

HRI kinome 0.97 0.86 0.97 M-005007-00 27102 NM_014413

HSA250839 kinome 1.01 0.99 0.98 M-004618-00 55351 NM_018401

HSMDPKIN kinome 1.65 0.64 0.79 M-007691-00 55561 XM_290516

HUNK kinome 0.90 0.31 0.74 M-004214-01 30811 NM_014586

ICAP-1A kinome 1.21 1.21 0.82 M-011927-00 9270 NM_004763

ICK kinome 0.64 0.77 0.95 M-004811-01 22858 NM_014920

IGF1R kinome 1.13 0.49 0.78 M-003012-04 3480 NM_000875

IHPK1 kinome 0.87 0.99 0.92 M-006737-01 9807 NM_153273

IHPK2 kinome 0.84 0.96 0.95 M-006738-00 51447 NM_016291

IHPK3 kinome 1.03 0.89 0.88 M-006739-00 117283 NM_054111

IKBKAP kinome 1.07 0.88 0.95 M-009371-00 8518 NM_003640

IKBKB kinome 0.76 0.79 0.95 M-003503-00 3551 XM_032491

IKBKE kinome 1.03 1.46 0.96 M-003723-02 9641 NM_014002

IL2 kinome 1.11 0.97 1.05 M-007967-00 3558 NM_000586

ILK kinome 1.37 0.82 0.80 M-004499-00 3611 NM_004517

ILKAP kinome 1.14 0.77 0.95 M-010260-00 80895 NM_030768

IMPK kinome 0.84 0.98 0.95 M-006740-01 253430 NM_152230

INSR motility 0.42 1.28 0.90 M-003014-01 3643 NM_000208

INSR kinome 0.62 0.93 0.87 M-003014-01 3643 NM_000208

INSR motility 0.93 1.08 0.90 M-003014-01 3643 NM_000208

INSRR kinome 0.99 0.89 0.87 M-005332-00 3645 NM_014215

IRAK1 kinome 0.65 0.68 0.87 M-004760-02 3654 NM_001569

IRAK2 kinome 1.24 1.06 1.14 M-004761-00 3656 NM_001570

IRAK3 kinome 0.66 0.37 0.74 M-004762-00 11213 NM_007199

IRS1 kinome 0.85 0.71 0.85 M-003015-01 3667 NM_005544

ITK kinome 0.87 1.06 0.97 M-003144-01 3702 NM_005546

ITPK1 kinome 1.16 0.69 0.72 M-006741-00 3705 NM_014216

ITPKA kinome 0.91 1.14 0.92 M-006742-01 3706 NM_002220

ITPKB kinome 1.38 0.89 1.02 M-006743-01 3707 NM_002221

ITPKC kinome 1.04 1.11 0.89 M-006744-01 80271 NM_025194

JAK1 kinome 0.94 0.84 0.95 M-003145-01 3716 NM_002227

JAK2 kinome 0.87 1.12 0.97 M-003146-02 3717 NM_004972

JAK2 motility 0.87 1.35 1.13 M-003146-02 3717 NM_004972

JAK2 motility 1.11 1.39 1.21 M-003146-02 3717 NM_004972

JAK3 kinome 0.96 0.78 0.90 M-003147-01 3718 NM_000215

JIK kinome 0.79 0.87 1.04 M-004844-01 51347 NM_016281

KDR kinome 0.79 0.78 0.95 M-003148-01 3791 NM_002253

KHK kinome 1.00 1.20 1.02 M-006745-01 3795 NM_000221

KIAA0551 kinome 1.14 0.70 0.74 M-004542-02 23043 XM_039796

KIAA0561 kinome 0.78 0.99 0.94 M-004046-00 23031 XM_038150

KIAA0999 kinome 1.75 0.56 0.87 M-004779-02 23387 NM_025164

KIAA1361 kinome 0.52 0.78 0.81 M-004846-01 57551 XM_290796

KIAA1399 kinome 0.69 0.83 0.85 M-023172-00 57574 XM_046685

KIAA1639 kinome 0.94 1.08 1.01 M-005336-00 57729 XM_290923

KIAA1765 kinome 0.61 1.19 1.06 M-005337-00 85443 XM_047355

KIAA1804 kinome 0.66 1.01 1.02 M-004063-00 84451 NM_032435

KIAA1811 kinome 0.86 0.97 0.99 M-004619-01 84446 NM_032430

KIAA1883 kinome 0.61 0.70 0.84 M-005338-01 114783 XM_055866

KIDINS220 motorpr 1.02 0.90 1.00 M-022984-00 57498 XM_291015

KIF11 motorpr 1.43 0.32 0.35 M-003317-01 3832 NM_004523

KIF12 motorpr 1.04 1.49 0.74 M-008252-01 113220 NM_138424

KIF13A motorpr 0.53 1.52 0.97 M-008257-01 63971 NM_022113

KIF13B motorpr 0.97 0.98 1.14 M-004963-00 23303 NM_015254

KIF13B kinome 1.15 0.91 0.95 M-004963-00 23303 NM_015254

KIF14 motorpr 0.85 0.46 0.88 M-003319-00 9928 NM_014875

KIF15 motorpr 1.23 1.44 0.85 M-004960-00 56992 NM_020242

KIF17 motorpr 0.19 1.55 0.68 M-022067-01 57576 NM_020816

KIF18A motorpr 1.24 1.05 0.87 M-006849-00 81930 NM_031217

KIF1A motorpr 1.40 0.96 0.84 M-010236-00 547 NM_004321

KIF1B motorpr 0.63 0.62 0.88 M-009317-00 23095 NM_015074

KIF1C motorpr 0.20 0.56 0.59 M-010354-01 10749 NM_006612

KIF2 motorpr 1.08 0.90 0.99 M-004959-01 3796 NM_004520

KIF20A motorpr 0.96 0.82 0.85 M-004957-01 10112 NM_005733

KIF21A motorpr 0.86 0.78 0.88 M-004969-00 55605 NM_017641

KIF21B motorpr 0.61 1.15 1.05 M-027602-00 23046 XM_371332

KIF22 motorpr 1.59 1.04 0.87 M-004962-00 3835 NM_007317

KIF23 motorpr 0.80 0.78 0.51 M-004956-01 9493 NM_004856

KIF24 motorpr 0.96 0.75 0.93 M-032566-00 55265 AK001795

KIF25 motorpr 0.78 1.06 1.00 M-010082-00 3834 NM_005355

KIF26A motorpr 1.56 0.61 0.76 M-022010-01 26153 XM_050278

KIF27 motorpr 0.80 1.08 1.01 M-008723-00 55582 NM_017576

KIF2B motorpr 0.27 0.67 1.02 M-008345-01 84643 NM_032559

KIF2C motorpr 0.94 1.46 0.85 M-004955-01 11004 NM_006845

KIF3A motorpr 0.62 1.06 1.22 M-004964-02 11127 NM_007054

KIF3B motorpr 0.82 0.75 0.85 M-009595-01 9371 NM_004798

KIF3C motorpr 0.86 0.75 0.92 M-009469-01 3797 NM_002254

KIF4A motorpr 1.21 1.26 1.03 M-004961-00 24137 NM_012310

KIF5A motorpr 0.91 0.85 1.12 M-008559-01 3798 NM_004984

KIF5B motorpr 1.06 0.87 1.02 M-008867-00 3799 NM_004521

KIF5C motorpr 0.81 1.18 1.18 M-019811-00 3800 XM_377774

KIF9 motorpr 0.47 0.30 0.78 M-004967-00 64147 NM_022342

KIFC1 motorpr 1.13 1.57 1.01 M-004958-01 3833 XM_371813

KIFC2 motorpr 0.54 0.72 1.04 M-008786-00 90990 NM_145754

KIFC3 motorpr 0.94 0.80 0.95 M-008338-01 3801 NM_005550

KIS kinome 0.89 0.83 0.92 M-003981-01 127933 NM_144624

KIT kinome 1.00 0.83 1.00 M-003150-01 3815 NM_000222

KIT motility 1.02 0.97 1.06 M-003150-01 3815 NM_000222

KLC2 motorpr 1.11 0.27 0.84 M-014218-01 64837 NM_022822

KLC2L motorpr 1.28 0.97 1.09 M-016063-00 147700 NM_177417

KNS2 motorpr 0.75 0.64 0.95 M-019482-01 3831 NM_005552

KNSL8 motorpr 0.76 0.88 1.09 M-015510-00 89953 NM_138343

KPI2 kinome 1.20 0.74 0.95 M-003149-01 22853 NM_014916

KRAS2 kinome 0.53 0.70 0.83 M-005069-00 3845 NM_004985

KSR2 kinome 1.02 0.95 0.86 M-005322-00 283455 NM_173598

LAK kinome 1.39 0.75 0.96 M-005009-00 80216 NM_025144

LATS1 kinome 1.04 0.97 0.89 M-004632-00 9113 NM_004690

LATS2 kinome 0.85 1.07 0.97 M-003865-00 26524 NM_014572

LCK kinome 0.83 0.61 0.87 M-003151-01 3932 NM_005356

LCP2 kinome 1.03 1.01 1.07 M-012120-00 3937 NM_005565

LIM kinome 0.52 0.40 0.88 M-006930-00 10611 NM_006457

LIMK1 kinome 0.72 0.80 0.75 M-007730-00 3984 NM_002314

LIMK1 motility 1.20 0.70 0.96 M-007730-01 3984 NM_002314

LIMK2 kinome 0.91 0.78 0.96 M-003311-02 3985 NM_005569

LOC115704 kinome 1.18 0.80 0.96 M-015901-00 115704 NM_145245

LOC146909 motorpr 0.78 0.91 1.08 M-010460-00 146909 XM_085634

LOC149420 kinome 0.77 0.95 1.01 M-005011-00 149420 NM_152835

LOC200383 motorpr 0.88 0.84 1.10 M-016271-00 200383 NM_145299

LOC340371 kinome 0.65 0.83 0.97 M-005340-00 340371 NM_178564

LOC374654 motorpr 0.57 1.10 0.65 M-022322-00 374654 NM_198525

LOC645357 motorpr 1.09 0.78 0.92 M-034981-00 645357 XM_928398

LOC647083 motorpr 1.11 0.49 0.77 M-032982-00 647083 XM_930094

LOC91807 kinome 1.15 1.05 0.86 M-005342-01 91807 NM_182493

LTK kinome 0.88 0.93 1.04 M-003152-01 4058 NM_002344

LYK5 kinome 0.90 0.68 0.85 M-005343-00 92335 NM_153335

LYN kinome 1.15 0.38 0.79 M-003153-03 4067 NM_002350

M-RIP motility 1.25 0.93 0.93 M-014102-00 23164 NM_015134

MADH7 kinome 0.79 0.72 0.94 M-020068-00 4092 NM_005904

MAGI-3 kinome 0.64 1.12 0.95 M-009453-00 260425 NM_020965

MAK kinome 0.57 0.71 0.95 M-004813-01 4117 NM_005906

MALT1 kinome 0.78 0.74 0.91 M-005936-00 10892 NM_006785

MAP1LC3B kinome 1.10 0.88 0.88 M-012846-00 81631 NM_022818

MAP2K1 kinome 1.11 0.98 0.91 M-003571-00 5604 NM_002755

MAP2K1IP1 kinome 0.69 0.94 0.98 M-003572-02 8649 NM_021970

MAP2K2 kinome 0.95 0.95 1.06 M-003573-03 5605 NM_030662

MAP2K3 kinome 0.84 0.98 0.86 M-003509-01 5606 NM_002756

MAP2K3 motility 0.89 0.82 0.80 M-003509-01 5606 NM_002756

MAP2K3 motility 1.15 0.69 0.82 M-003509-01 5606 NM_002756

MAP2K4 kinome 0.67 0.88 0.94 M-003574-02 6416 NM_003010

MAP2K5 kinome 1.23 0.86 0.87 M-003966-03 5607 NM_002757

MAP2K6 motility 0.61 0.76 1.04 M-003967-00 5608 NM_002758

MAP2K6 motility 0.90 0.57 1.00 M-003967-00 5608 NM_002758

MAP2K6 kinome 0.97 0.44 0.86 M-003967-00 5608 NM_002758

MAP2K7 kinome 0.43 0.32 0.65 M-004016-01 5609 NM_145185

MAP3K1 kinome 1.25 0.75 0.90 M-003575-01 4214 XM_042066

MAP3K10 motility 0.77 0.42 0.57 M-003576-01 4294 NM_002446

MAP3K10 kinome 1.07 0.86 0.91 M-003576-01 4294 NM_002446

MAP3K10 motility 1.09 0.74 0.71 M-003576-01 4294 NM_002446

MAP3K11 kinome 0.97 0.85 0.81 M-003577-01 4296 NM_002419

MAP3K12 kinome 1.02 0.92 0.84 M-003312-02 7786 NM_006301

MAP3K12 motility 1.15 1.01 0.82 M-003312-02 7786 NM_006301

MAP3K12 motility 1.25 0.92 0.85 M-003312-02 7786 NM_006301

MAP3K13 kinome 0.83 1.06 1.02 M-003579-01 9175 NM_004721

MAP3K14 kinome 1.05 0.64 0.82 M-003580-02 9020 NM_003954

MAP3K2 kinome 1.28 0.82 0.94 M-003582-01 10746 NM_006609

MAP3K3 kinome 0.96 0.47 0.91 M-003301-02 4215 NM_002401

MAP3K4 kinome 1.11 1.00 0.91 M-003789-02 4216 NM_005922

MAP3K5 kinome 0.80 1.02 0.97 M-003584-00 4217 NM_005923

MAP3K6 kinome 0.81 1.59 0.92 M-003969-00 9064 NM_004672

MAP3K7 kinome 0.88 1.09 0.88 M-003790-04 6885 NM_003188

MAP3K8 kinome 1.02 0.93 0.99 M-003511-00 1326 NM_005204

MAP3K9 kinome 0.93 0.71 0.90 M-003585-01 4293 XM_027237

MAP4K1 kinome 1.08 1.94 0.70 M-003586-01 11184 NM_007181

MAP4K2 kinome 0.79 0.49 0.84 M-003587-01 5871 NM_004579

MAP4K3 kinome 0.77 0.81 0.93 M-003588-00 8491 NM_003618

MAP4K4 kinome 0.99 0.63 0.94 M-003971-02 9448 NM_004834

MAP4K5 kinome 1.00 0.89 1.02 M-003589-01 11183 NM_006575

MAPK1 kinome 0.39 0.63 0.82 M-003555-02 5594 NM_002745

MAPK10 kinome 0.80 0.86 1.01 M-004324-00 5602 NM_002753

MAPK11 kinome 1.70 0.40 0.71 M-003972-03 5600 NM_002751

MAPK12 kinome 0.72 0.60 0.82 M-003590-00 6300 NM_002969

MAPK12 motility 0.91 0.57 0.79 M-003590-00 6300 NM_002969

MAPK12 motility 0.94 0.58 0.80 M-003590-00 6300 NM_002969

MAPK13 kinome 1.12 0.88 0.97 M-003591-02 5603 NM_002754

MAPK14 kinome 0.86 1.30 1.07 M-003512-03 1432 NM_001315

MAPK3 kinome 0.73 0.82 1.00 M-003592-02 5595 NM_002746

MAPK4 kinome 0.82 1.10 1.07 M-003593-00 5596 NM_002747

MAPK6 kinome 0.84 1.07 0.90 M-003594-01 5597 NM_002748

MAPK7 kinome 1.61 0.88 0.89 M-003513-02 5598 NM_002749

MAPK8 kinome 1.14 1.10 1.03 M-003514-01 5599 NM_002750

MAPK8IP1 kinome 0.60 1.01 1.10 M-003595-00 9479 NM_005456

MAPK8IP2 kinome 0.89 0.50 0.76 M-012462-00 23542 NM_012324

MAPK8IP3 kinome 0.96 1.04 0.90 M-003596-01 23162 NM_015133

MAPK9 kinome 1.05 1.00 1.01 M-003505-02 5601 NM_002752

MAPKAPK2 kinome 0.90 0.71 0.90 M-003516-01 9261 NM_004759

MAPKAPK3 kinome 1.24 0.87 1.05 M-005014-00 7867 NM_004635

MAPKAPK5 kinome 0.72 1.54 0.94 M-005015-00 8550 NM_003668

MARCKS kinome 0.91 1.53 0.87 M-004772-00 4082 NM_002356

MARK1 kinome 0.69 0.55 0.87 M-004259-02 4139 NM_018650

MARK1 motility 0.89 0.60 0.96 M-004259-02 4139 NM_018650

MARK1 motility 0.96 0.41 0.94 M-004259-02 4139 NM_018650

MARK2 kinome 0.90 0.90 0.87 M-004260-01 2011 NM_004954

MARK3 kinome 1.24 0.23 0.63 M-003517-03 4140 NM_002376

MARKL1 kinome 0.54 0.81 0.90 M-005345-02 57787 NM_031417

MAST205 kinome 1.18 0.91 0.89 M-004633-00 23139 NM_015112

MATK kinome 0.86 1.20 1.05 M-003154-03 4145 NM_002378

MBIP kinome 0.87 1.21 1.07 M-021396-00 51562 NM_016586

MCL1 kinome 1.17 1.05 0.87 M-004501-02 4170 NM_021960

MDM2 motility 0.58 1.06 0.92 M-003279-02 4193 NM_002392

MDM2 kinome 1.02 1.12 0.86 M-003279-02 4193 NM_002392

MELK kinome 0.99 0.90 1.00 M-004029-00 9833 NM_014791

MERTK kinome 1.27 0.85 0.93 M-003155-01 10461 NM_006343

MET kinome 0.30 1.20 1.09 M-003156-01 4233 NM_000245

MGC16169 kinome 0.80 0.82 0.85 M-005346-00 93627 NM_033115

MGC22688 kinome 0.97 1.07 1.06 M-004634-00 202374 NM_145001

MGC26597 kinome 0.85 0.81 0.81 M-004783-00 206426 NM_152700

MGC3248 motorpr 1.72 1.09 0.98 M-014901-00 84516 NM_032486

MGC33182 kinome 0.69 1.23 1.01 M-004681-01 122011 NM_145203

MGC42105 kinome 0.93 0.94 1.15 M-005016-00 167359 NM_153361

MGC43306 kinome 0.91 0.72 0.87 M-005347-00 169436 XM_291304

MGC45428 kinome 1.11 0.73 0.80 M-005017-00 166614 NM_152619

MGC46424 kinome 0.96 0.74 0.95 M-008770-00 138429 NM_173492

MGC4796 kinome 0.88 0.88 1.00 M-005348-00 83931 NM_032017

MGC5601 kinome 0.63 1.07 0.87 M-007684-00 80724 NM_025247

MGC8407 kinome 0.95 0.70 0.88 M-005349-00 79012 NM_024046

MIDORI kinome 1.02 1.22 0.78 M-005350-01 57538 NM_020778

MINK kinome 1.10 0.27 0.91 M-004861-02 50488 NM_015716

MKNK1 kinome 1.00 0.70 0.95 M-004879-01 8569 NM_003684

MKNK2 kinome 0.92 0.98 0.98 M-004908-00 2872 NM_017572

MLC1SA motorpr 0.72 0.74 1.03 M-013090-00 140465 NM_002475

MOS kinome 0.92 0.81 1.01 M-003859-02 4342 NM_005372

MPHOSPH1 motorpr 0.68 0.92 1.09 M-003318-01 9585 NM_016195

MPP1 kinome 1.22 1.25 0.95 M-010252-00 4354 NM_002436

MPP2 kinome 0.62 0.76 0.95 M-009729-00 4355 NM_005374

MPP3 kinome 0.80 1.28 0.97 M-010612-00 4356 NM_001932

MPZL1 kinome 0.87 0.80 1.05 M-015738-00 9019 NM_003953

MRC2 kinome 0.79 0.73 0.80 M-020064-00 9902 NM_006039

MRCL3 motorpr 0.62 0.96 0.67 M-021498-00 10627 NM_006471

MRLC2 motorpr 0.38 1.47 0.88 M-018116-00 103910 NM_033546

MSN kinome 0.78 0.86 0.93 M-011732-00 4478 NM_002444

MST1R kinome 1.06 0.50 0.69 M-003157-02 4486 NM_002447

MST4 kinome 1.27 0.98 1.04 M-003753-01 51765 NM_016542

MUSK kinome 0.71 0.36 0.87 M-003158-01 4593 NM_005592

MVD kinome 0.76 0.43 0.85 M-006748-00 4597 NM_002461

MVK kinome 0.84 0.78 0.88 M-006749-00 4598 NM_000431

MYC kinome 0.94 0.69 0.75 M-003282-01 4609 NM_002467

MYH1 motorpr 0.73 0.58 0.96 M-013486-00 4619 NM_005963

MYH10 motorpr 1.39 1.41 1.16 M-023017-00 4628 NM_005964

MYH11 motorpr 0.22 0.36 0.85 M-011737-00 4629 NM_002474

MYH11 motorpr 0.88 0.13 0.89 M-011737-00 4629 NM_002474

MYH13 motorpr 1.36 0.77 0.91 M-012621-00 8735 NM_003802

MYH14 motorpr 0.87 0.42 0.63 M-027149-00 79784 NM_024729

MYH15 motorpr 0.51 1.03 0.99 M-023571-00 22989 XM_036988

MYH2 motorpr 0.87 0.66 0.95 M-021252-00 4620 NM_017534

MYH3 motorpr 0.85 1.00 0.88 M-012644-00 4621 NM_002470

MYH4 motorpr 0.57 0.97 0.95 M-021250-00 4622 NM_017533

MYH6 motorpr 0.84 1.14 0.89 M-012645-00 4624 NM_002471

MYH7 motorpr 1.36 0.73 0.77 M-011086-00 4625 NM_000257

MYH7B motorpr 0.59 0.43 0.82 M-031354-00 57644 XM_371398

MYH8 motorpr 1.03 0.84 1.03 M-012646-00 4626 NM_002472

MYH9 motorpr 0.97 1.72 0.60 M-007668-00 4627 NM_002473

MYL5 motorpr 1.16 0.85 0.78 M-011739-00 4636 NM_002477

MYL6 motorpr 0.70 1.24 0.98 M-013091-00 4637 NM_021019

MYL6 motorpr 0.74 1.13 0.97 M-013091-00 4637 NM_021019

MYL7 motorpr 0.38 1.07 0.67 M-021478-00 58498 NM_021223

MYL9 motorpr 1.24 0.93 0.90 M-019044-00 10398 NM_006097

MYLC2PL motorpr 0.96 0.61 0.93 M-015564-00 93408 NM_138403

MYLIP motorpr 0.41 0.78 0.94 M-006976-00 29116 NM_013262

MYLK motorpr 1.25 0.95 0.63 M-005351-04 4638 NM_005965

MYLK kinome 1.54 0.91 0.83 M-005351-01 4638 NM_005965

MYLK2 motorpr 0.81 1.08 0.83 M-005352-02 85366 NM_033118

MYLK2 kinome 1.04 1.02 0.73 M-005352-01 85366 NM_033118

MYO10 motorpr 1.39 0.90 1.01 M-007217-00 4651 NM_012334

MYO15A motorpr 0.52 0.75 0.84 M-013160-00 51168 NM_016239

MYO18A motorpr 1.00 0.60 0.87 M-031755-00 399687 NM_078471

MYO18B motorpr 1.46 0.92 1.03 M-015302-00 84700 NM_032608

MYO1A motorpr 1.00 0.83 0.97 M-008765-00 4640 NM_005379

MYO1B motorpr 0.83 0.83 1.02 M-023110-00 4430 NM_012223

MYO1C motorpr 0.63 0.84 1.01 M-015121-00 4641 NM_033375

MYO1D motorpr 1.16 0.64 0.87 M-023316-00 4642 XM_050041

MYO1E motorpr 0.45 0.52 0.92 M-019919-00 4643 NM_004998

MYO1F motorpr 1.02 0.68 1.01 M-017712-00 4542 NM_012335

MYO1G motorpr 0.98 0.81 0.91 M-025326-00 64005 XM_291223

MYO3A kinome 1.12 1.28 0.87 M-004862-00 53904 NM_017433

MYO3A motorpr 1.65 0.96 0.75 M-004862-00 53904 NM_017433

MYO3B kinome 0.77 1.16 1.03 M-004863-00 140469 NM_138995

MYO3B motorpr 0.86 1.18 1.03 M-004863-00 140469 NM_138995

MYO5A motorpr 0.83 1.12 0.93 M-019321-00 4644 NM_000259

MYO5B motorpr 0.73 0.91 1.05 M-023431-00 4645 XM_371116

MYO5C motorpr 0.60 1.23 1.11 M-031960-00 55930 NM_018728

MYO6 motorpr 0.99 0.46 1.10 M-006355-00 4646 NM_004999

MYO7A motorpr 1.20 1.27 0.94 M-019330-00 4647 NM_000260

MYO7B motorpr 1.55 0.96 1.10 M-022928-00 4648 XM_291001

MYO9A motorpr 1.00 0.81 0.87 M-006539-00 4649 NM_006901

MYO9B motorpr 1.02 0.89 0.81 M-009535-00 4650 NM_004145

MYR8 motorpr 0.90 0.74 0.97 M-021586-00 23026 XM_028522

NAGK kinome 1.04 0.35 0.57 M-006750-00 55577 NM_017567

NBEA kinome 1.02 0.90 1.06 M-015419-00 26960 NM_015678

NBP kinome 1.00 0.42 0.72 M-006751-00 80347 NM_025233

NCK1 motility 0.91 0.95 0.97 M-006354-01 4690 NM_006153

NCK2 motility 0.57 0.38 0.94 M-019547-00 8440 NM_003581

NCK2 motility 0.73 0.44 0.81 M-019547-00 8440 NM_003581

NEK1 kinome 1.16 0.83 0.88 M-004864-00 4750 XM_291107

NEK11 kinome 0.78 0.49 0.91 M-004865-01 79858 NM_024800

NEK2 kinome 0.82 0.62 1.05 M-004090-02 4751 NM_002497

NEK3 kinome 1.14 1.01 0.88 M-004867-00 4752 NM_002498

NEK4 kinome 0.97 1.32 0.98 M-003519-01 6787 NM_003157

NEK6 motility 0.77 0.63 1.14 M-004166-01 10783 NM_014397

NEK6 kinome 1.09 0.80 1.02 M-004166-00 10783 NM_014397

NEK7 kinome 0.89 0.47 0.79 M-003795-01 140609 NM_133494

NEK8 kinome 0.98 0.85 0.65 M-004866-00 284086 NM_178170

NEK9 kinome 0.85 0.90 0.81 M-004869-01 91754 NM_033116

NF2 kinome 0.85 0.57 0.91 M-003917-00 4771 NM_000268

NLK kinome 0.51 0.95 0.74 M-004763-01 51701 NM_016231

NME1 kinome 0.92 1.33 0.85 M-006821-00 4830 NM_000269

NME2 kinome 0.94 1.14 0.84 M-005102-00 4831 NM_002512

NME3 kinome 0.83 0.85 0.92 M-006753-00 4832 NM_002513

NME4 kinome 0.97 1.02 0.85 M-006494-00 4833 NM_005009

NME5 kinome 1.19 0.91 0.99 M-006754-00 8382 NM_003551

NME6 kinome 0.89 0.86 0.92 M-006755-01 10201 NM_005793

NME7 kinome 0.83 0.96 0.99 M-006756-01 29922 NM_013330

NPR1 kinome 0.74 0.87 1.03 M-005354-00 4881 NM_000906

NPR2 kinome 0.79 1.13 0.91 M-005355-00 4882 NM_000907

NRAS kinome 1.09 0.86 0.97 M-003919-00 4893 NM_002524

NRBP kinome 0.62 0.98 1.03 M-005356-00 29959 NM_013392

NRG3 kinome 0.88 0.80 0.95 M-026286-00 10718 XM_166086

NTRK1 kinome 0.78 1.11 0.87 M-003159-01 4914 NM_002529

NTRK2 kinome 0.78 1.06 1.00 M-003160-01 4915 NM_006180

NTRK3 kinome 0.97 0.99 1.02 M-003161-01 4916 NM_002530

NYD-SP25 kinome 0.96 1.04 0.99 M-007733-00 89882 NM_033516

OSBPL1A motorpr 0.76 0.94 0.92 M-008350-01 114876 NM_018030

OSR1 kinome 1.16 0.97 0.99 M-004870-00 9943 NM_005109

P15RS kinome 0.82 1.03 1.05 M-007734-00 55197 NM_018170

PACE-1 kinome 0.55 0.53 0.82 M-005357-00 57147 NM_020423

PACSIN1 kinome 0.99 0.71 0.79 M-007735-00 29993 NM_020804

PAG kinome 1.07 1.08 0.95 M-012956-00 55824 NM_018440

PAK1 motility 0.72 0.76 1.01 M-003521-03 5058 NM_002576

PAK1 kinome 0.78 0.70 0.88 M-003521-03 5058 NM_002576

PAK1 motility 1.07 0.58 1.05 M-003521-03 5058 NM_002576

PAK2 motility 0.55 0.76 0.88 M-003597-02 5062 NM_002577

PAK2 kinome 0.80 0.78 0.77 M-003597-02 5062 NM_002577

PAK2 motility 0.83 0.65 0.90 M-003597-02 5062 NM_002577

PAK3 kinome 0.82 0.85 1.01 M-003614-00 5063 NM_002578

PAK4 kinome 0.90 1.01 0.91 M-003615-01 10298 NM_005884

PAK6 kinome 0.76 0.83 1.00 M-004338-01 56924 NM_020168

PAK7 kinome 0.97 0.98 0.91 M-003973-02 57144 NM_020341

PANK1 kinome 0.91 0.76 0.96 M-004057-00 53354 NM_138316

PANK3 kinome 0.92 0.90 0.99 M-006758-00 79646 NM_024594

PANK4 kinome 1.18 1.02 1.00 M-006759-00 55229 NM_018216

PAPSS1 kinome 1.19 0.61 0.86 M-007736-00 9061 NM_005443

PAPSS2 kinome 1.21 0.89 0.94 M-006760-00 9060 NM_004670

PASK kinome 1.13 1.06 0.94 M-005018-01 23178 NM_015148

PCK1 kinome 0.89 0.59 0.68 M-006796-00 5105 NM_002591

PCK2 kinome 0.89 0.69 0.87 M-006797-00 5106 NM_004563

PCTK1 kinome 1.05 0.76 0.77 M-004313-00 5127 NM_006201

PCTK2 kinome 1.28 0.73 0.91 M-004835-01 5128 NM_002595

PCTK3 kinome 0.85 0.85 0.86 M-004836-02 5129 NM_002596

PDGFRA kinome 1.18 0.80 0.84 M-003162-02 5156 NM_006206

PDGFRB kinome 0.62 0.69 0.95 M-003163-02 5159 NM_002609

PDK1 kinome 0.78 0.73 0.80 M-005019-00 5163 NM_002610

PDK2 kinome 1.09 0.51 0.92 M-005020-00 5164 NM_002611

PDK3 kinome 0.84 0.91 1.02 M-005021-00 5165 NM_005391

PDK4 kinome 0.83 0.59 0.84 M-019425-00 5166 NM_002612

PDPK1 kinome 1.08 0.53 0.94 M-003017-01 5170 NM_002613

PDXK kinome 1.11 1.17 1.05 M-005070-00 8566 NM_003681

PEA15 motility 1.17 0.91 1.12 M-010553-01 8682 NM_003768

PEA15 motility 1.28 0.88 1.01 M-010553-01 8682 NM_003768

PFKFB1 kinome 0.89 1.02 1.00 M-006761-00 5207 NM_002625

PFKFB2 kinome 1.06 0.91 0.90 M-006762-01 5208 NM_006212

PFKFB3 kinome 0.80 0.95 0.96 M-006763-00 5209 NM_004566

PFKFB4 kinome 1.04 1.02 1.03 M-006764-00 5210 NM_004567

PFKL kinome 1.15 1.13 0.92 M-006822-00 5211 NM_002626

PFKM kinome 1.10 0.93 1.03 M-006765-00 5213 NM_000289

PFKP kinome 0.80 0.65 0.75 M-010253-00 5214 NM_002627

PFTK1 kinome 0.75 1.06 1.00 M-004837-00 5218 NM_012395

PGK1 kinome 0.77 1.01 1.02 M-006767-01 5230 NM_000291

PGK2 kinome 0.89 0.98 0.98 M-006768-01 5232 NM_138733

PHKA1 kinome 1.09 0.93 0.97 M-019682-00 5255 NM_002637

PHKA2 kinome 0.99 0.81 0.95 M-007669-00 5256 NM_000292

PHKG1 kinome 0.66 1.01 0.94 M-005023-01 5260 NM_006213

PHKG2 kinome 0.67 0.74 0.84 M-004881-00 5261 NM_000294

PI4K2B kinome 0.99 1.02 1.00 M-006769-00 55300 NM_018323

PI4KII kinome 1.12 0.97 1.00 M-006770-00 55361 NM_018425

PIK3C2A kinome 1.19 0.87 0.99 M-006771-00 5286 NM_002645

PIK3C2B kinome 0.82 0.74 0.94 M-006772-01 5287 NM_002646

PIK3C2G kinome 0.69 1.11 0.94 M-006773-00 5288 NM_004570

PIK3C3 kinome 1.04 0.92 0.94 M-005250-00 5289 NM_002647

PIK3CA kinome 1.05 0.68 0.81 M-003018-01 5290 NM_006218

PIK3CB kinome 0.89 0.63 0.86 M-003019-01 5291 NM_006219

PIK3CD kinome 0.89 1.23 0.86 M-006775-01 5293 NM_005026

PIK3CG kinome 0.81 0.43 0.82 M-005274-00 5294 NM_002649

PIK3R1 kinome 1.08 0.75 0.83 M-003020-02 5295 NM_181504

PIK3R2 kinome 0.92 0.49 0.99 M-003021-01 5296 NM_005027

PIK3R3 kinome 1.04 0.71 0.99 M-019546-00 8503 NM_003629

PIK3R4 kinome 1.36 0.59 0.89 M-005025-00 30849 NM_014602

PIK4CA kinome 1.87 0.80 0.84 M-006776-03 5297 NM_002650

PIK4CB kinome 0.78 0.97 0.89 M-006777-02 5298 NM_002651

PIM1 kinome 1.11 0.97 0.91 M-003923-00 5292 NM_002648

PIM2 kinome 0.87 0.78 0.73 M-005359-00 11040 NM_006875

PINK1 kinome 0.79 0.72 0.83 M-004030-00 65018 NM_032409

PIP5K1A kinome 1.43 0.48 0.82 M-004780-02 8394 NM_003557

PIP5K2A kinome 0.81 0.98 0.93 M-006778-00 5305 NM_005028

PIP5K2B kinome 1.14 0.74 0.86 M-006779-01 8396 NM_003559

PIP5K2C kinome 0.73 0.87 0.96 M-004535-00 79837 NM_024779

PITPNM3 kinome 1.12 0.95 0.82 M-014699-00 83394 NM_031220

PKE kinome 0.77 1.14 0.94 M-004615-00 282974 NM_173575

PKIA kinome 0.72 0.66 0.89 M-012321-00 5569 NM_006823

PKIB kinome 1.62 0.82 1.05 M-008224-00 5570 NM_032471

PKLR kinome 1.21 0.91 0.94 M-006780-00 5313 NM_000298

PKM2 kinome 0.93 0.91 0.80 M-006781-00 5315 NM_002654

PKMYT1 kinome 0.77 0.83 0.84 M-005026-02 9088 NM_004203

PKNBETA kinome 0.56 0.81 0.86 M-004647-00 29941 NM_013355

PLK kinome 0.97 0.54 0.76 M-003290-01 5347 NM_005030

PMSCL2 kinome 0.67 1.10 0.96 M-010904-00 5394 NM_002685

PMVK kinome 0.74 0.48 0.93 M-006782-00 10654 NM_006556

PNKP kinome 0.53 0.34 0.72 M-006783-01 11284 NM_007254

POLE motility 1.12 0.65 0.87 M-020132-00 5426 NM_006231

PPP1R1B kinome 0.96 1.05 0.94 M-012745-00 84152 NM_032192

PPP2CA kinome 0.97 0.72 1.04 M-003598-00 5515 NM_002715

PPP2CB kinome 0.87 0.84 0.95 M-003599-00 5516 NM_004156

PPP4C kinome 0.73 1.00 0.95 M-008486-00 5531 NM_002720

PRKAA1 kinome 0.89 0.87 1.00 M-005027-00 5562 NM_006251

PRKAA2 kinome 0.70 1.19 0.95 M-005361-00 5563 NM_006252

PRKACA kinome 0.83 0.87 0.95 M-004649-00 5566 NM_002730

PRKACB kinome 0.95 1.00 1.01 M-004650-00 5567 NM_002731

PRKACG kinome 0.97 1.00 1.07 M-004651-01 5568 NM_002732

PRKAG1 kinome 1.00 0.99 0.98 M-009056-00 5571 NM_002733

PRKAG3 kinome 0.29 0.93 0.71 M-009859-00 53632 NM_017431

PRKAR1A kinome 1.48 0.94 0.97 M-007670-00 5573 NM_002734

PRKAR2A kinome 1.27 0.91 0.99 M-007671-00 5576 NM_004157

PRKAR2B kinome 0.58 0.95 0.87 M-007673-00 5577 NM_002736

PRKCA kinome 1.09 1.04 0.91 M-003523-02 5578 NM_002737

PRKCABP kinome 0.79 1.18 0.91 M-020124-00 9463 NM_012407

PRKCB1 kinome 1.03 1.08 0.92 M-003758-03 5579 NM_002738

PRKCD kinome 1.66 0.81 0.89 M-003524-01 5580 NM_006254

PRKCE kinome 0.96 0.70 0.92 M-004653-00 5581 NM_005400

PRKCG kinome 0.71 0.70 0.97 M-004654-00 5582 NM_002739

PRKCH kinome 0.87 1.05 0.98 M-004655-01 5583 NM_006255

PRKCI kinome 1.37 1.11 1.09 M-004656-00 5584 NM_002740

PRKCL1 kinome 1.16 0.66 0.95 M-004175-02 5585 NM_002741

PRKCL2 kinome 1.11 0.60 0.90 M-004612-02 5586 NM_006256

PRKCM kinome 1.04 0.87 0.95 M-005028-00 5587 NM_002742

PRKCM motility 1.17 0.78 1.04 M-005028-00 5587 NM_002742

PRKCN kinome 0.25 1.05 0.90 M-005029-00 23683 NM_005813

PRKCQ kinome 0.92 0.79 0.90 M-003525-01 5588 NM_006257

PRKCSH kinome 1.06 0.68 0.96 M-010618-00 5589 NM_002743

PRKCZ motility 0.49 0.94 1.06 M-003526-02 5590 NM_002744

PRKCZ kinome 0.74 0.95 0.98 M-003526-02 5590 NM_002744

PRKD2 kinome 1.10 0.94 0.94 M-004197-01 25865 NM_016457

PRKDC kinome 0.93 0.68 0.88 M-005030-00 5591 NM_006904

PRKG1 kinome 0.60 0.61 0.94 M-004658-02 5592 NM_006258

PRKG2 kinome 0.99 1.04 1.05 M-004659-00 5593 NM_006259

PRKR kinome 0.85 0.99 1.02 M-003527-00 5610 NM_002759

PRKRA kinome 1.12 0.78 0.97 M-006426-00 8575 NM_003690

PRKWNK1 kinome 0.66 0.88 0.84 M-005362-00 65125 NM_018979

PRKWNK2 kinome 0.64 1.01 0.97 M-005363-01 65268 NM_006648

PRKWNK3 kinome 0.74 0.98 1.06 M-005364-01 65267 NM_020922

PRKWNK4 kinome 1.15 0.65 0.92 M-005031-00 65266 NM_032387

PRKX kinome 1.02 0.97 0.93 M-004660-01 5613 NM_005044

PRKY kinome 1.15 0.92 1.01 M-004661-02 5616 NM_002760

PRPF4B kinome 0.36 0.76 0.81 M-004074-03 8899 NM_003913

PRPS1 kinome 0.73 0.89 0.96 M-006784-00 5631 NM_002764

PRPS1L1 kinome 1.40 0.84 0.96 M-006804-00 221823 NM_175886

PRPS2 kinome 0.92 0.85 1.06 M-004877-00 5634 NM_002765

PRPSAP1 kinome 1.16 1.16 0.85 M-006794-01 5635 NM_002766

PRPSAP2 kinome 0.62 0.68 0.80 M-006795-00 5636 NM_002767

PRSS25 motility 0.98 1.30 0.97 M-006014-02 27429 NM_013247

PRSS25 motility 1.47 1.13 1.00 M-006014-02 27429 NM_013247

PSK kinome 0.65 0.78 0.88 M-004171-02 51677 NM_016151

PSKH1 kinome 1.19 0.95 0.88 M-005365-00 5681 NM_006742

PSKH2 kinome 0.80 0.58 0.92 M-005366-00 85481 NM_033126

PTEN kinome 0.67 1.06 1.07 M-003023-01 5728 NM_000314

PTK2 motility 0.78 0.67 0.74 M-003164-02 5747 NM_005607

PTK2 motility 0.87 0.94 0.78 M-003164-02 5747 NM_005607

PTK2 kinome 0.92 0.62 0.78 M-003164-01 5747 NM_005607

PTK2B kinome 1.33 0.87 0.79 M-003165-03 2185 NM_004103

PTK2B motility 1.44 1.16 0.84 M-003165-03 2185 NM_004103

PTK2B motility 1.55 0.99 0.81 M-003165-03 2185 NM_004103

PTK6 kinome 1.31 0.73 0.91 M-003166-01 5753 NM_005975

PTK7 kinome 0.82 1.08 0.88 M-003167-01 5754 NM_002821

PTK9 kinome 0.65 0.87 0.96 M-003168-02 5756 NM_002822

PTK9L kinome 0.87 0.60 0.87 M-003169-01 11344 NM_007284

PTPN5 kinome 1.23 1.14 0.79 M-003600-01 84867 NM_032781

PTPRG kinome 0.87 0.86 0.97 M-008069-00 5793 NM_002841

PTPRJ kinome 1.15 0.58 0.95 M-008476-00 5795 NM_002843

PTPRR kinome 0.94 0.92 1.04 M-004017-00 5801 NM_002849

PTPRT kinome 1.01 0.81 0.90 M-008072-00 11122 NM_007050

PXK kinome 1.16 1.02 0.92 M-005367-00 54899 NM_017771

PXN motility 0.53 0.76 0.83 M-005163-00 5829 NM_002859

PXN kinome 0.54 0.48 0.83 M-005163-00 5829 NM_002859

PXN motility 0.67 0.57 0.84 M-005163-00 5829 NM_002859

PYCS kinome 1.18 0.47 0.56 M-006785-00 5832 NM_002860

RAB24 motorpr 1.00 0.99 0.97 M-008828-00 53917 NM_130781

RAB4A motorpr 0.94 1.37 1.15 M-008539-01 5867 NM_004578

RAB5A motorpr 0.52 0.68 0.84 M-004009-00 5868 NM_004162

RAB5B motorpr 1.47 1.02 0.99 M-004010-01 5869 NM_002868

RAB5C motorpr 0.88 0.93 1.00 M-004011-01 5878 NM_004583

RAB7 motorpr 1.38 0.97 0.87 M-010388-00 7879 NM_004637

RAB7B motorpr 1.38 1.15 1.01 M-018225-00 338382 NM_177403

RAB7L1 motorpr 1.50 1.00 0.85 M-010556-01 8934 NM_003929

RAB8A motorpr 0.62 1.24 0.98 M-003905-00 4218 NM_005370

RAB8B motorpr 0.68 0.61 1.03 M-008744-00 51762 NM_016530

RAC1 kinome 0.85 0.30 0.83 M-003560-02 5879 NM_006908

RAC1 motility 1.32 0.35 0.79 M-003560-02 5879 NM_018890

RAC2 motility 0.75 0.63 0.99 M-007741-00 5880 NM_002872

RAC2 kinome 1.18 0.68 0.86 M-007741-00 5880 NM_002872

RAC3 motility 0.90 0.80 0.85 M-008836-01 5881 NM_005052

RAC3 kinome 1.16 0.92 0.91 M-008836-00 5881 NM_005052

RAF1 kinome 1.04 0.66 0.80 M-003601-00 5894 NM_002880

RAGE kinome 0.92 0.63 0.99 M-004838-00 5891 NM_014226

RALA kinome 0.78 0.74 0.80 M-009235-00 5898 NM_005402

RALB kinome 0.89 0.73 0.81 M-008403-00 5899 NM_002881

RALGDS kinome 1.09 0.92 0.84 M-005193-00 5900 NM_006266

RASGRF2 kinome 0.88 1.03 0.93 M-024516-00 5924 NM_006909

RBSK kinome 0.95 0.89 0.89 M-006786-00 64080 NM_022128

RDX kinome 1.59 0.45 0.90 M-011762-00 5962 NM_002906

RET kinome 0.67 0.88 0.93 M-003170-01 5979 NM_000323

RFP kinome 1.44 1.02 0.92 M-006552-00 5987 NM_006510

RGL1 kinome 0.72 0.97 0.96 M-008387-00 23179 NM_015149

RGL2 kinome 1.04 0.69 0.95 M-009321-00 5863 NM_004761

RHEB kinome 1.14 0.56 0.73 M-009692-00 6009 NM_005614

RHOA motility 0.63 1.10 0.89 M-003860-00 387 NM_001664

RHOA kinome 0.69 0.96 0.90 M-003860-00 387 NM_001664

RHOA motility 0.82 1.05 0.98 M-003860-00 387 NM_001664

RHOB kinome 0.75 0.70 0.99 M-008395-00 388 NM_004040

RHOB motility 1.05 1.16 1.09 M-008395-01 388 NM_004040

RHOB motility 1.17 0.98 1.12 M-008395-01 388 NM_004040

RHOBTB1 motility 0.73 0.84 0.99 M-009389-00 9886 NM_014836

RHOBTB1 motility 1.00 0.76 1.02 M-009389-00 9886 NM_014836

RHOBTB2 motility 0.74 1.07 0.98 M-009252-00 23221 NM_015178

RHOBTB2 motility 0.87 1.08 0.97 M-009252-00 23221 NM_015178

RHOBTB3 motility 1.17 1.16 1.04 M-020480-01 22836 NM_014899

RHOC kinome 1.00 0.83 0.93 M-008555-00 389 NM_175744

RHOC motility 1.07 0.80 0.87 M-008555-00 389 NM_175744

RHOC motility 1.25 0.68 0.81 M-008555-00 389 NM_175744

RHOD motility 1.15 1.07 0.91 M-008940-00 29984 NM_014578

RHOD motility 1.19 1.02 0.89 M-008940-00 29984 NM_014578

RHOF motility 0.88 1.05 0.99 M-008316-00 54509 NM_019034

RHOG motility 0.76 0.63 0.97 M-008995-00 391 NM_001665

RHOH motility 0.75 0.76 1.02 M-008804-00 399 NM_004310

RHOJ motility 0.92 1.03 1.05 M-010367-00 57381 NM_020663

RHOJ motility 1.24 0.83 0.81 M-010367-00 57381 NM_020663

RHOK kinome 0.86 0.89 0.95 M-004662-00 6011 NM_002929

RHOQ motility 0.63 1.48 1.02 M-009943-00 23433 NM_012249

RHOQ motility 0.85 1.62 1.10 M-009943-00 23433 NM_012249

RHOT1 motility 1.05 0.88 0.96 M-010365-00 55288 NM_018307

RHOT2 motility 0.57 1.16 1.05 M-008340-00 89941 NM_138769

RHOU motility 0.85 0.86 1.00 M-009882-00 58480 NM_021205

RHOU motility 0.88 1.03 1.06 M-009882-00 58480 NM_021205

RHOV motility 0.87 1.38 1.01 M-006374-00 171177 NM_133639

RHOV motility 0.96 1.30 0.97 M-006374-00 171177 NM_133639

RILP motorpr 0.48 1.29 0.82 M-008787-00 83547 NM_031430

RIOK1 kinome 1.16 0.52 0.74 M-005368-01 83732 NM_031480

RIOK3 kinome 1.21 1.53 1.00 M-005040-00 8780 NM_003831

RIPK1 kinome 1.00 0.88 0.87 M-004445-01 8737 NM_003804

RIPK2 kinome 1.26 0.84 0.88 M-003602-00 8767 NM_003821

RIPK3 kinome 0.78 0.81 0.80 M-003534-00 11035 NM_006871

RNASEL kinome 0.66 0.84 0.85 M-005032-00 6041 NM_021133

RND1 motility 1.20 1.01 1.05 M-008929-00 27289 NM_014470

ROCK1 kinome 0.46 0.97 0.92 M-003536-01 6093 NM_005406

ROCK2 kinome 0.89 1.07 1.07 M-004610-01 9475 NM_004850

ROR1 kinome 1.18 1.10 0.89 M-003171-01 4919 NM_005012

ROR2 kinome 1.32 0.66 0.99 M-003172-01 4920 NM_004560

ROS1 kinome 0.94 0.70 0.98 M-003173-01 6098 NM_002944

RP2 kinome 1.18 0.96 1.00 M-012350-00 6102 NM_006915

RPS6KA1 kinome 0.78 1.01 0.82 M-003025-03 6195 NM_002953

RPS6KA2 kinome 0.79 0.87 0.96 M-004663-00 6196 NM_021135

RPS6KA3 kinome 0.52 0.91 0.99 M-003026-01 6197 NM_004586

RPS6KA4 kinome 0.44 0.91 0.87 M-004664-00 8986 NM_003942

RPS6KA5 kinome 0.82 0.97 1.03 M-004665-00 9252 NM_004755

RPS6KA6 kinome 0.79 1.14 1.05 M-004670-00 27330 NM_014496

RPS6KB1 kinome 0.72 0.88 0.81 M-003616-02 6198 NM_003161

RPS6KB2 kinome 1.20 0.80 1.02 M-004671-00 6199 NM_003952

RPS6KC1 kinome 1.28 0.92 1.09 M-005371-00 26750 NM_012424

RPS6KL1 kinome 1.06 0.58 0.79 M-005372-00 83694 NM_031464

RYK kinome 1.03 0.93 0.98 M-003174-02 6259 NM_002958

SAST kinome 1.07 0.67 0.94 M-004672-01 22983 XM_032034

SCAP1 kinome 1.14 0.95 0.96 M-011505-00 8631 NM_003726

SCYL1 kinome 0.36 1.22 1.04 M-005373-00 57410 NM_020680

SEPHS1 kinome 1.01 0.79 0.82 M-007737-00 22929 NM_012247

SGK kinome 0.93 1.26 1.00 M-003027-03 6446 NM_005627

SGK2 kinome 1.07 1.13 0.97 M-004673-00 10110 NM_016276

SGKL kinome 1.20 1.05 1.02 M-004162-00 23678 NM_013257

SH3MD1 motility 0.95 0.90 1.07 M-006657-01 9644 XM_374831

SHC1 kinome 0.95 0.81 0.92 M-018841-00 6464 NM_003029

SIK2 kinome 0.52 0.58 0.89 M-004778-02 23235 NM_015191

SLK kinome 1.24 1.23 0.89 M-003850-01 9748 NM_014720

SMG1 kinome 1.15 1.05 0.83 M-005033-00 23049 NM_014006

SNARK kinome 0.67 0.71 0.89 M-005374-01 81788 NM_030952

SNF1LK kinome 0.75 0.88 0.98 M-003959-04 150094 NM_173354

SNK kinome 0.80 1.14 0.96 M-003325-03 10769 NM_006622

SNRK kinome 0.91 1.08 1.06 M-004322-00 54861 NM_017719

SOCS1 kinome 0.79 0.73 0.89 M-011511-00 8651 NM_003745

SOCS5 kinome 1.12 1.22 1.01 M-017374-00 9655 NM_014011

SPA17 kinome 1.25 0.83 0.88 M-007677-00 53340 NM_017425

SPEC2 kinome 0.88 1.01 0.97 M-020826-00 56990 NM_020240

SPHK1 kinome 0.83 0.96 0.93 M-004172-02 8877 NM_021972

SPHK2 kinome 1.30 0.78 0.84 M-004831-00 56848 NM_020126

SPRED1 kinome 0.97 1.18 1.02 M-016638-00 161742 NM_152594

SPRED2 kinome 1.16 0.97 0.90 M-018590-00 200734 NM_181784

SPS2 kinome 0.57 1.14 0.94 M-007738-00 22928 NM_012248

SQSTM1 kinome 0.96 1.05 0.90 M-010230-00 8878 NM_003900

SRC kinome 0.92 0.79 0.86 M-003175-03 6714 NM_005417

SRMS kinome 0.85 0.72 0.77 M-005376-00 6725 NM_080823

SRPK1 kinome 0.76 1.10 1.09 M-003982-01 6732 NM_003137

SRPK2 kinome 1.06 1.08 0.95 M-004839-01 6733 NM_182691

SSH3BP1 kinome 0.81 0.85 0.93 M-007290-00 10006 NM_005470

SSTK kinome 1.03 0.64 0.90 M-005034-00 83983 NM_032037

STK10 kinome 1.45 0.67 0.85 M-004168-01 6793 NM_005990

STK11 kinome 0.62 0.85 0.88 M-005035-00 6794 NM_000455

STK16 kinome 1.20 1.07 1.02 M-004054-00 8576 NM_003691

STK17A kinome 0.82 0.62 0.91 M-005377-00 9263 NM_004760

STK17B kinome 0.83 0.90 0.93 M-004051-01 9262 NM_004226

STK18 kinome 0.77 0.95 0.80 M-005036-00 10733 NM_014264

STK19 kinome 0.93 1.24 0.93 M-005378-00 8859 NM_004197

STK22B kinome 0.78 0.71 0.94 M-005379-01 23617 NM_053006

STK22C kinome 1.21 0.87 1.01 M-004050-01 81629 NM_052841

STK22D kinome 0.93 1.13 1.01 M-005038-01 83942 NM_032028

STK23 kinome 0.82 0.76 1.00 M-004840-01 26576 NM_014370

STK24 kinome 1.17 0.52 0.81 M-004872-01 8428 NM_003576

STK25 kinome 1.43 0.81 0.91 M-004873-00 10494 NM_006374

STK29 kinome 0.79 0.70 1.00 M-005381-01 9024 NM_003957

STK3 kinome 0.21 0.95 1.05 M-004874-00 6788 NM_006281

STK31 kinome 0.76 0.77 0.92 M-005382-00 56164 NM_031414

STK33 kinome 0.94 0.79 0.92 M-005383-01 65975 NM_030906

STK35 kinome 0.72 0.70 0.78 M-005384-00 140901 NM_080836

STK36 kinome 0.91 0.77 0.85 M-005039-00 27148 NM_015690

STK38 kinome 0.46 0.57 0.94 M-004674-00 11329 NM_007271

STK38L kinome 0.91 1.11 0.92 M-004875-00 23012 NM_015000

STK39 kinome 1.30 0.76 1.01 M-004157-01 27347 NM_013233

STK4 kinome 0.87 0.36 0.69 M-003545-09 6789 NM_006282

STK6 motility 0.54 0.46 0.81 M-003545-09 6790 NM_003600

STK6 motility 0.68 0.46 0.82 M-003545-09 6790 NM_003600

STK6 kinome 0.85 1.08 0.97 M-003313-02 6790 NM_003600

STMN1 kinome 0.74 0.76 0.93 M-005152-00 3925 NM_005563

STMN2 kinome 0.99 1.09 0.98 M-019342-00 11075 NM_007029

STMN3 kinome 0.72 0.73 0.86 M-013600-00 50861 NM_015894

SYK kinome 1.43 0.20 0.59 M-003176-03 6850 NM_003177

TAF1 kinome 0.73 0.82 0.60 M-005041-00 6872 NM_004606

TAF1L kinome 0.86 0.93 0.79 M-005385-01 138474 NM_153809

TAO1 kinome 1.19 0.59 0.76 M-004171-02 9344 NM_004783

TBK1 kinome 0.63 0.70 0.75 M-003788-02 29110 NM_013254

TCTE1L motorpr 0.95 1.22 1.03 M-019957-00 6990 NM_006520

TCTEL1 motorpr 1.02 0.94 0.81 M-019964-00 6993 NM_006519

TEC kinome 0.97 0.73 1.03 M-003177-02 7006 NM_003215

TEK kinome 1.19 0.79 0.98 M-003178-02 7010 NM_000459

TESK1 kinome 0.54 0.96 0.98 M-005043-00 7016 NM_006285

TESK2 kinome 1.19 0.89 0.77 M-005044-00 10420 NM_007170

TEX14 kinome 1.56 0.67 0.83 M-005386-00 56155 NM_031272

TGFBR1 kinome 0.71 0.53 0.82 M-003929-01 7046 NM_004612

TGFBR2 kinome 1.04 0.86 0.99 M-003930-00 7048 NM_003242

TIAM1 kinome 1.11 0.46 0.78 M-003932-00 7074 NM_003253

TIE kinome 1.20 1.05 0.91 M-003179-01 7075 NM_005424

TJP2 kinome 1.16 0.89 0.80 M-009932-00 9414 NM_004817

TK1 kinome 1.00 1.11 0.90 M-006787-00 7083 NM_003258

TK2 kinome 1.07 0.66 0.89 M-006788-02 7084 NM_004614

TLK1 kinome 0.84 0.95 1.15 M-004174-00 9874 NM_012290

TLK2 kinome 1.05 1.17 0.99 M-005389-02 11011 NM_006852

TLN1 motility 1.33 0.35 0.99 M-012949-00 7094 NM_006289

TLR1 kinome 0.76 0.99 0.97 M-008086-00 7096 NM_003263

TLR3 kinome 0.86 0.99 0.78 M-007745-00 7098 NM_003265

TLR4 kinome 1.16 0.49 0.48 M-008088-00 7099 NM_003266

TLR6 kinome 0.47 0.22 0.60 M-005156-00 10333 NM_006068

TNFRSF10A kinome 0.62 0.74 1.00 M-008090-00 8797 NM_003844

TNK1 kinome 1.13 0.59 0.94 M-003180-01 8711 NM_003985

TOPK kinome 1.14 1.10 0.96 M-005390-00 55872 NM_018492

TP53 kinome 0.70 0.55 1.00 M-003329-01 7157 NM_000546

TP73 kinome 0.89 0.69 0.97 M-003331-01 7161 NM_005427

TPK1 kinome 0.45 0.47 0.70 M-006789-00 27010 NM_022445

TRAD kinome 0.69 0.76 0.68 M-005045-00 11139 NM_007064

TRB2 kinome 1.14 0.26 0.63 M-005391-00 28951 NM_021643

TRIM kinome 1.41 0.42 0.68 M-020821-00 50852 NM_016388

TRIO kinome 1.23 0.98 0.82 M-005047-00 7204 NM_007118

TRPM6 kinome 0.75 1.05 0.99 M-005048-01 140803 NM_017662

TRPM7 kinome 1.06 0.37 0.77 M-005393-00 54822 NM_017672

TSC1 kinome 0.70 1.07 0.95 M-003028-01 7248 NM_000368

TSC2 kinome 0.55 1.09 0.96 M-003029-01 7249 NM_000548

TSKS kinome 1.18 0.52 0.84 M-013077-00 60385 NM_021733

TTBK1 kinome 1.06 0.83 0.96 M-004680-01 84630 XM_166453

TTBK2 kinome 0.89 0.75 0.79 M-004682-00 146057 NM_173500

TTK kinome 0.58 0.63 0.52 M-004105-00 7272 NM_003318

TTN kinome 0.72 0.75 1.14 M-005395-00 7273 NM_133378

TTN kinome 0.93 1.00 1.04 M-005395-00 7273 NM_133378

TXK kinome 0.85 0.83 0.92 M-003181-01 7294 NM_003328

TXNDC3 kinome 0.97 1.08 0.91 M-006791-00 51314 NM_016616

TYK2 kinome 0.62 0.35 0.61 M-003182-01 7297 NM_003331

TYRO3 kinome 0.77 0.25 0.69 M-003183-01 7301 NM_006293

UCK1 kinome 1.26 0.72 0.91 M-004062-00 83549 NM_031432

UGP2 kinome 0.99 1.20 0.97 M-007739-01 7360 NM_006759

ULK1 kinome 0.99 1.31 1.02 M-005049-00 8408 NM_003565

ULK2 kinome 0.85 0.80 0.94 M-005396-01 9706 NM_014683

UMP-CMPK kinome 0.43 0.76 0.88 M-004059-00 51727 NM_016308

UMPK kinome 0.69 0.77 0.86 M-005077-00 7371 NM_012474

URKL1 kinome 0.71 0.93 1.01 M-006792-00 54963 NM_017859

VCL motility 0.68 0.80 0.91 M-009288-00 7414 NM_003373

VCL motility 0.72 0.78 1.00 M-009288-00 7414 NM_003373

VIL2 kinome 0.94 0.88 0.86 M-017370-01 7430 NM_003379

VRK1 kinome 1.08 1.14 0.95 M-004683-01 7443 NM_003384

VRK2 kinome 1.03 0.83 0.84 M-004684-01 7444 NM_006296

VRK3 kinome 1.00 0.75 0.77 M-005397-00 51231 NM_016440

WASF1 motility 0.54 1.23 1.17 M-011557-00 8936 NM_003931

WASF1 motility 0.72 1.14 1.13 M-011557-00 8936 NM_003931

WASF2 motility 1.48 0.48 0.57 M-012141-00 10163 NM_006990

WASF2 motility 1.49 0.61 0.71 M-012141-00 10163 NM_006990

WASL motility 1.00 0.92 1.16 M-006444-01 8976 NM_003941

WASL motility 1.15 0.90 1.18 M-006444-01 8976 NM_003941

WEE1 kinome 1.01 0.68 0.74 M-005050-00 7465 NM_003390

WIF1 kinome 0.79 1.14 0.89 M-012386-00 11197 NM_007191

XYLB kinome 0.80 0.67 1.02 M-006793-00 9942 NM_005108

YES1 kinome 1.03 0.74 0.96 M-003184-02 7525 NM_005433

YWHAH kinome 0.83 1.38 0.83 M-010626-00 7533 NM_003405

YWHAQ kinome 0.61 0.90 0.89 M-012329-00 10971 NM_006826

ZAK kinome 0.99 1.05 1.08 M-005068-00 51776 NM_016653

ZAP70 kinome 1.00 0.70 0.67 M-005398-01 7535 NM_001079

ZW10 motility 0.79 0.75 0.89 M-003948-02 9183 NM_004724

ZYX motility 0.78 1.21 0.99 M-016734-00 7791 NM_003461

ZYX motility 1.12 1.18 0.99 M-016734-00 7791 NM_003461