FIGURE 5.

N-WASP is involved in Gas7-induced membrane protrusions in Neuro-2a cells. A, Neuro-2a cells co-transfected with Gas7 (red) and either ECFP (control, green) or N-WASP mutants (ECFP-CA or HA/N-WASPΔcof) (green) were immunostained, and nuclei were counterstained with Hoechst 33258 (cyan). In the lower right panel, the arrow shows successful extension of membrane protrusion; the arrowhead shows a lack of extension. Scale bar, 10 μm. B, shown is the fractions of cells in A with membrane protrusions. Data are presented as the mean ± S.E. of three independent determinations, each counting at least 100 cells. Statistically significant decreases of the fraction of cells that extend processes compared with the control group (ECFP/Gas7 co-transfection) are indicated (paired Student's t test; *, p < 0.05; **, p < 0.01). C, extracts from COS-7 cells transfected with Gas7/Myc alone (negative control) or in combination with either wild-type HA/N-WASP (positive control) or HA/N-WASPΔcof were subjected to co-IP to assay the interaction between HA-N-WASPΔcof and Gas7. Gas7 was detected in complex with both N-WASPΔcof and N-WASP.