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. 2009 Apr 24;284(17):11622–11636. doi: 10.1074/jbc.M805940200

FIGURE 4.

FIGURE 4.

N-WASP-induced filopodial formation requires and synergizes with Toca-1. A, N1E115 cells were transfected with N-WASP alone, N-WASP with Toca-1 mutants, and N-WASP with Toca-1 (a). All transfections were carried out with GFP-actin. Panels show GFP and DIC images with a merge. The bar chart shows a statistical analysis of the experiments (b). Neurites and filopodial formation were scored as described under “Materials and Methods.” Toca-1 F-BAR domain and Cdc42 binding mutants were as described in the legend to Fig. 2. Three independent experiments were performed (with n = 10-15). Data shown are mean ± S.D. Bar = 10 μm. B, Toca-1 was KD using RNAi and the affect of N-WASP on filopodial formation examined. a, Toca-1 protein levels with control RNAi and Toca-1 RNAi. b, left panel, N-WASP transfection with control RNAi. Right panel, N-WASP transfection with Toca-1 RNAi. Lower panel shows tracing of filopodia in red. c, statistical analysis of experiments similar to those presented in b of single cells. Data shown are mean ± S.D. (n = 20).