N-WASP KD does not affect lamellipod or filopod formation. (A) Representative images of cells treated with scrambled or N-WASP siRNA. Cells were fixed and stained with Rhodamine phalloidin at 0 and 3 min after EGF stimulation. Bar, 10 μm. (B) Quantification of lamellipod extension (cell area). Cell areas were measured at 0 and 3 min after EGF stimulation. The graph shows the fold increase in area after EGF stimulation of scrambled siRNA and N-WASP siRNA. A total of 41 cells pooled from three independent experiments are shown. (C) Quantification of filopod formation. A total of 30 cells from three independent experiments are shown. (D) Western blot of cells treated with N-WASP–specific or scrambled (Scr) siRNA and blotted with α–N-WASP and α–β actin antibodies. Error bars indicate ±SEM.