Fig. 3

YAP1 promotes EMT and upregulates DCLK1 by repressing miR-124-3p and miR-188-5p. A, B Western blot analysis of the expression levels of YAP1, DCLK1 or Vimentin in HCT116 and SW620 cells transfected with two different YAP1 siRNAs. β-actin was used as loading control. Statistical analysis was performed compared with siNC group. C, D Western blot analysis of YAP1, DCLK1 or Vimentin in HCT116 and SW620 cells transfected with YAP1 siRNA and then rescued with transfection of Flag-YAP-5SA plasmid. Flag tag was used to indicate the overexpression of YAP-5SA. E, F RT-qPCR analysis of DCLK1 and Vimentin at mRNA levels and concurrent expression levels of miR-124-3p and miR-188-5p in HCT116 and SW620 cells transfected with YAP1 siRNAs. Statistical analysis was performed compared with siNC group. G RT-qPCR analysis of expression levels of miR-124-3p and miR-188-5p in HCT116 cells transfected with YAP1 siRNA and Flag-YAP-5SA plasmid compared with siNC and vector control respectively. H, I Western blot analysis of YAP1, DCLK1 or Vimentin in CXCR7-overexpressing HCT116 and SW620 cells transfected with YAP1 siRNA and siNC. The vector control CRC cells were transfected with siNC as controls. Bars are means ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 (n = 3)