WAVE2 is required for lateral membrane motility. (A) Co-staining for DAAM1, WAVE2, and F-actin in EpH4 cells treated with control or DAAM1 siRNA. Enlarged views of the boxed regions are shown at the right. DAAM1 and WAVE2 showed distinct distributions, and WAVE2 overlapped with F-actin. (B) Immunostaining for E-cadherin in EpH4 cells treated with DAAM1 and/or WAVE2 siRNAs. WAVE2 KD tended to cause the sharpening of E-cadherin signals at the basal edges of LCs in both control and DAAM1-depleted cells. (A and B) Arrowheads point to the basal edges of the junctions. (C) Quantification of the tilting extent of LCs. (D) DAAM1-depleted cells were treated with control or WAVE2 siRNAs for 1 d and then labeled with CMTPX. After mixing them with nonlabeled shControl cells, they were cultured on collagen gels for 2 d. Protrusion in DAAM1-depleted cells was suppressed by WAVE2 KD. Arrows indicate representative protrusions. Bars: (A [left], B, and D) 10 µm; (A, right) 5 µm. (E) Quantification of protrusion number and length in the experiment shown in D. (C and E) Histograms represent the mean of three experiments. Error bars indicate SD. **, P < 0.01; ***, P < 0.001; n.s., not significant.