Fig. 4
Disruption of ETV6 leads to TWIST1-dependent malignant phenotypes in prostate cancer cells. (a) Western blot assay of PC3 cells with stable expression of ETV6 followed by transient transfection of a TWIST1-expressing vector. EV and ctrl, control vectors of ETV6 and TWIST1, respectively. (b, c) PC3 cells with a combination of ETV6 and TWIST1 expressions as in panel a were analyzed by migration (b) or invasion (c) assays. Selected images are shown on the right. (d) Quantification of TWIST1 mRNA and the Western blot assay in derivative cell lines of DU145. A cell line with stable ETV6-knockdown (shETV6) was generated by a lentiviral approach. shLacZ, control. (e) Survival analysis of mice challenged with derivatives of the DU145 cell line. (f) Selected images of brain tissues from mice treated with prostate cancer cells as in panel e. The metastatic tumor is distinguished from normal brain tissues by a dashed line and is labeled with an arrow. (g) Western blot assay of LNCaP cells with a combination of ETV6- and TWIST1-knockdown by specific siRNA (siETV6, siETV6 + siTWSIT1). scr, control siRNA. (h) Migration and invasion assays of LNCaP cells with a combination of ETV6 and TWIST1 knockdown. (i) Representative images of results from panel h. Data are presented as the mean ± SEM, n = 3. *p < 0.05, **p < 0.01, *** p < 0.001, ****p < 0.0001