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Tspan14. Tspan12. CD9. -. GFP- Tspan :. +. +. +. +. HA-ADAM10:. A. i.p : GFP- Tspan blot: HA-ADAM10. **. 125. 80. 80. 100. HA-ADAM10 quantitation from GFP i.p . Relative ADAM10 level. 75. 58. 58. 58. 50. 25. 0. B. whole cell lysate blot: HA-ADAM10. **. 60. 40. - PowerPoint PPT Presentation
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46
2530
i.p: GFP-Tspan
blot: GFP-Tspan
C
% m
atur
e A
DA
M10 60
40
20
0
HA-ADAM10 quantitation
from whole cell lysate
58
**
whole cell lysate
blot: HA-ADAM108058
B
Rel
ativ
eA
DA
M10
leve
l
HA-ADAM10 quantitation
from GFP i.p.
1007550250
125
i.p: GFP-Tspan
blot: HA-ADAM108058
A
Tspa
n14
-
+ + + +GFP-Tspan:
HA-ADAM10:
CD
9
Tspa
n12
**
Supplementary Figure III. The non-TspanC8 tetraspanin Tspan12 does not interact with or promote ADAM10 maturation. The experiment was performed and presented as described in Figure 2A-C, with the exception that GFP-tagged mouse tetraspanins were used instead of FLAG-tagged tetraspanins. The GFP-tagged mouse forms of non-TspanC8 tetraspanins CD9 and Tspan12 were compared to Tspan14 in their capacity to (A) co-immunoprecipitate with, and (B) promote maturation of, bovine HA-tagged ADAM10 (n=5; **p<0.01 compared to all other data; error bars represent standard errors of the mean). The GFP immunoprecipitation was performed with GFP-Trap beads and confirmation of similar Tspan12 and Tspan14 expression was obtained by (C) anti-GFP western blotting.