Effect of WNT10B knockdown on EMT, metalloproteases and stemness gene expression and the stem-like cell population in PC3 cells. A, PC3 WNT10B shRNA cells showed an increased expression of E-cadherin and reduced expression of N-cadherin, vimentin, MMP9, NANOG, and SOX2 compared with control shRNA-PC3 cells. *P < .05 vs control cells, N = 3. Bars represent the mean ± SEM. B, Flow cytometry utilizing a Hoechst exclusion assay was used to assess effect of WNT10B knockdown in PC3 cells on the stem-like cell side population. Control shRNA and WNT10B shRNA-PC3 cells were stained with 5 μg/mL of Hoechst33342 either in the absence (left) or presence (right) of verapamil hydrochloride 15 minutes before fluorescence-activated cell sorting. The graph represents the percentage side-population cells (gated population) in control shRNA and WNT10B shRNA-PC3 cells. Knockdown of WNT10B significantly abrogated the stem-like cell side population. *P < .001 vs control cells, N = 4. Bars represent the mean ± SEM. EMT, epithelial to mesenchymal transition; shRNA, short hairpin