Fig. 5: Knockdown of Sox9 imitates miR-133b-mediated phenotypes in MDA-MB-231.

a Detection of endogenous WAVE2 and Sox9 expression by western blot at 48 h after transfection of 100 nM siWAVE2 or siSox9, respectively, in MDA-MB-231 cells. β-Actin serves as an internal control. b, c Effects of transfection of siWAVE2 or siSox9 on the proliferation of MDA-MB-231 cells. b Morphology of MDA-MB-231 cells at day 4 and day 6 post transfection of siWAVE2 or siSox9 (scale bars, 100 μm). c Proliferation curves of MDA-MB-231 cells transfected with siWAVE2 or siSox9. Triplicate wells from each cohort were counted for 6 days. d Influence of siWAVE2 or siSox9 on colony formation of MDA-MB-231 cells. Left panel: representative images by colony formation assay. Right panel: the number of clones were counted for each well of six-well plates and shown in the y axis. The results were reproducible in three independent experiments. e Transwell migration and invasion assays of MDA-MB-231 cells transfected with 100 nM siWAVE2 or siSox9, respectively. The cell migration and invasion were quantitatively analyzed 18 h after seeding in Transwells. Data are presented as mean ± S.D. The symbol * denotes statistical difference (p < 0.05), whereas ** represents great significant difference (p < 0.01) in a two-tailed Student’s t-test. All experiments were repeated independently for three times