Figure 5.

EGF induces the colocalization of AMAP1 with PRKD2 via Rab5c. (A–F) MDA-MB-231 cells were cultured on collagen I–coated dishes, then transfected with Rab5c siRNA and pCMV-V5-PRKD2 one after another (A–C) or with pCMV-V5-PRKD2 and pcDNA3.1 HisC Rab5c simultaneously. Cells were serum-starved for 2 h, then treated (A and D) or left untreated (B and E) with 10 ng/ml EGF (5 min) before fixation. AMAP1, Xpress, and V5 epitopes were immunostained. In the merged image, V5-PRKD2 and AMAP1 are shown as green and red, respectively, in A and B; V5-PRKD2, Xpress-Rab5c, and AMAP1 are shown as green, red, and blue, respectively, in D and E. Colocalization indices of each protein were quantified, and the results represent mean ±SEM of >10 measurements (error bars; C and F). The rightmost panels in D and E show enlarged views of the boxed regions. Bars, 10 µm.