Figure 5.

EnaEVH1 Is Sufficient to Reduce Dia-Driven Filopodia

(A–C) D16 cells with GFP-DiaΔDAD alone (A), GFP-DiaΔDAD (B, B″, and B closeup) + mCh-EnaEVH1 (B′, B″, and B′ closeup), or GFP-DiaΔDAD+mCh-EnaEVH1^F77E (C). Arrows, cortical EnaEVH1.

(D) Mean filopodia number, DiaΔDAD alone (n = 27), DiaΔDAD+EnaEVH1 (n = 28), DiaΔDAD+EnaEVH1^F77E (n = 26), and GFP-DiaΔDAD+EnaProEVH2 (n = 29); error bars = 95% confidence interval.

(E) GST pull-down of DiaFH1 by GST, GST-EnaEVH1, or GST-EnaEVH1^F77E. S, supernatant; p, pellet. Bottom: Coomassie verifying equal load.

(F) Pyrene actin assembly with profilin and 10 nM DiaFH1FH2 (triangles), plus GST-EnaEVH1 (black diamonds) or GST-EnaEVH1^F77E (red diamonds).

(G) Time it takes 10 nM DiaFH1FH2 to stimulate 1/2 max steady-state pyrene fluorescence (maximum actin assembly) versus concentration of GST-EVH1 constructs.

See Figure S3.