Fig. 2

YAP was able to induced cell migration, invasion and focal adhesion in breast cancer cell lines. (a) Western blot verified the overexpression of YAP in MCF7 cells. EV: empty vector; o/e: overexpression. (b) Western blot verified the knockdown of YAP in MDA-MB-231 cells via a collection of siRNAs; siYAP-#2 and siYAP-#3 has relatively high knockdown efficiency, thus these two siRNAs were used in this research. (c, d) Transwell assay showing that overexpression of YAP induced cell migration and invasion ability in MCF7 cells. The experiment was performed in triplicate. **p < 0.01 by Student’s t-test. Scale bar: 100 μm. (e, f) Transwell assay showing that knockdown of YAP significantly inhibited cell migration and invasion ability in MDA-MB-231 cells. The experiment was performed in triplicate. ** p < 0.01 by ANOVA test. Scale bar: 100 μm. (g, h) Overexpression of YAP induced MCF7 cell adhesion to gelatin. The attached cells were stained with Wright’s-Giemsa and are shown in (g). The experiment was performed in triplicate. ** p < 0.01 by Student’s t-test. Scale bar: 100 μm. (i, j) Knockdown of YAP significantly inhibited MDA-MB-231 cell adhesion to gelatin. The attached cells were stained with Wright’s-Giemsa and are shown in (i). The experiment was performed in triplicate. ** p < 0.01 by Student’s t-test. Scale bar: 100 μm. (k) Overexpression of YAP induced focal adhesions in MCF7 cells. Focal adhesions were visualized by co-localization of paxilin (green) and F-actin (stained with phalloidin, red). Nuclei were counterstained with DAPI (blue). Scale bar: 20 μm. (l) Knockdown of YAP expression inhibited focal adhesions in MDA-MB-231 cells. Scale bar: 20 μm. (m) Quantification of the membrane-localized paxilin in (k). The experiment was performed in triplicate. ** p < 0.01 by Student’s t-test. (n) Quantification of the membrane-localized paxilin in (l). The experiment was performed in triplicate. ** p < 0.01 by ANOVA test