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. 2021 Mar 4;40(13):2407–2421. doi: 10.1038/s41388-021-01718-4

Fig. 4. Increase of YAP1 expression promotes cancer stemness and lipid metabolism in EnzaR cells.

Fig. 4

A RNA-seq results of YAP1 knockdown and its control in EnzaR cells were performed GSEA analyses. B Enzalutamide-resistant (EnzaR#2 and #3) and its parental cells (WT) were analyzed by flow cytometry. CD133 and CD44 double-positive cells were cells with cancer stemness (n = 3). C Enzalutamide-resistant (EnzaR#2 and #3) and its parental cells (WT) were performed sphere culture for 1 week. Results were presented as sphere number (n = 3). EnzaR#2 and its parental LNCaP cells were performed lipid staining by BODIPY 505/515 dye (D) and measured free fatty acid level by a commercial kit (E). E COUP-TFII (CII), YAP1, and BMI1 expression levels were detected by RT-qPCR in CD133− and CD133+ cells with stable knockdown of COUP-TFII or control construct sorted by CD133 magnetic beads. Results were normalized to 18s rRNA expression. F, G EnzaR#2 cells were treated with control (siNC), YAP1, or CII siRNA for 48 h. Then, those cells were trypsinized and performed sphere culture for 7 days. Representative pictures (F) and quantifications of sphere numbers (G) were shown. EnzaR#2 cells were individually or simultaneously knocked down YAP1 or COUP-TFII in EnzaR cells for 48 h and performed lipid staining (H) and measured free fatty acid level (I). Asterisk indicates p < 0.05 using one-way ANOVA test following by Dunnett analysis (B, C, G, I) and two-tailed Student’s t-test (E).