Supplemental figures for:

Essential Role for Rac1 in Cocaine-Induced Structural Plasticity of Nucleus

Accumbens Neurons

David M. Dietz, Haosheng Sun, Mary Kay Lobo, Michael E. Cahill, Benjamin

Chadwick, Virginia Gao, Ja Wook Koo, Michelle S. Mazei-Robison, Caroline

Dias, Ian Maze, Diane Damez-Werno, Karen C. Dietz, Kimberly N. Scobie,

Deveroux Ferguson, Daniel Christoffel, Yoko Ohnishi, Georgia E. Hodes, Yi

Zheng, Rachael L. Neve, Klaus M. Hahn, Scott J. Russo, and Eric J. Nestler

   

Nature Neuroscience: doi:10.1038/nn.3094

                                     Supplemental Figure 1. Rac1 mediated synaptic changes following chronic

cocaine.

Scheme depicting the mechanism underlying actin cytoskeleton reorganization

after repeated administration of cocaine. Compared to control conditions (a),

repeated cocaine (b), via repression of the GEF Tiam and induction of RacGap1,

primes NAc neurons for transient decreases in Rac1 activity in response to each

cocaine challenge, leading to dephosphorylation and increased activity of cofilin

and induction of thin spines.  

                     

Nature Neuroscience: doi:10.1038/nn.3094

       

Supplemental Figure 2: Full length blots for Fig. 1 (a-e)

Samples were loaded on a 4-15% SDS gel and then cut and blotted for (a) total

Rac1 following GTP-bound Rac1 pulldown 15 min or (b) 45 min following cocaine

treatments. Total Rac1 was probed using the same samples. (c) Samples were

loaded on a 4-15% SDS gel and then cut and blotted for phosphorylated cofilin

15 min and 4 hours following repeated cocaine. Total cofilin was used as a

loading control. Samples were loaded on a 4-15% SDS gel and then cut and

blotted for (d) Tiam1 or (e) Racgap1 twenty-four hours following cocaine

treatments. Tubulin was probed and used as a loading control.

             

Nature Neuroscience: doi:10.1038/nn.3094

                                 Supplemental Figure 3: Full length blots for Fig. 3 (a)

(a) Samples were loaded on a 4-15% SDS gel and then cut and blotted for

phosphorylated cofilin 15 min following blue light stimulation. Total cofilin was

used as a loading control.  

 

Nature Neuroscience: doi:10.1038/nn.3094