Breast Cancer Antiestrogen Resistance 3 (BCAR3) Promotes Cell Motility by Regulating Actin Cytoskeletal and Adhesion Remodeling in Invasive Breast Cancer Cells
Figure 7
RhoA effector signaling mediates adhesion length in BCAR3-depleted invasive breast cancer cells.
(A) BT549 cells transfected with siCtl or siB3-1 ± mDia1-targeted (siDia1) siRNA oligonucleotides were held in suspension for 90 minutes, plated onto 10 µg/ml fibronectin in the absence or presence of 20 µM Y-27632, and then processed for immunofluorescence as described in the methods. Adhesions were visualized using a vinculin antibody; insets show higher magnifications of peripheral adhesions. Scale bars = 15 µm. Similar results were obtained with paxillin staining (unpublished data). (B) Quantification of adhesion length in siCtl (black bars), siB3-1 (gray bars), siCtl+siDia1 (white bars), and siB3-1+siDia1 (hashed bars) treated cells in the absence or presence of Y-27632. Data represent the mean ± SEM of at least 343 adhesions from at least 13 cells from 2 separate experiments. ANOVA analysis confirmed all conditions were significantly different from one another (p<0.0001) except the following comparisons: siCtl-Y vs. siCtl+siDia1-Y (bars 1 vs. 5), siCtl+Y vs. siB3-1+Y (bars 2 vs. 4), and siCtl+siDia1+Y vs. siB3-1+siDia1+Y (bars 6 vs. 8).