Fig. 3

FOXA1 regulates enhancer activity to promote CYR61 expression. a TF binding to the CYR61 promoter, Enhancer1 and Enhancer3 were predicted by TRAP. b FOXA1 binding site predicted by TRAP. c CYR61 mRNA and eRNA levels decreased after treatment with FOXA1 siRNA for 72 h in RKO and SW480 cell lines. d CYR61 protein levels decreased after treatment with FOXA1 siRNA for 72 h in RKO and SW480 cell lines. e FOXA1 and H3K27ac enrichment after treatment with FOXA1 siRNA for 72 h in the RKO cell line, as assessed by ChIP-qPCR. f Relative crosslinking frequencies of CYR61 promoter/Enhancer1 and CYR61 promoter/Enhancer3 decreased after treatment with FOXA1 siRNA for 72 h in RKO cells, as assessed by 3C; significance determined by the paired-samples t-test. g Relative activity of promoter and enhancers after treatment of RKO cells with FOXA1 siRNA for 72 h, as assessed by relative luciferase reporter gene activity. (H) CYR61 mRNA and eRNA levels increased after transfection with GV219-mFOXA1 for 48 h in HCT116 and 72 h in SW480 cell lines. i CYR61 protein levels increased after transfection with GV219-mFOXA1 for 48 h in HCT116 and 72 h in SW480 cells. j FOXA1 and H3K27ac enrichment after transfection with GV219-mFOXA1 for 48 h in HCT116 cells, as assessed by ChIP-qPCR. k Relative crosslinking frequencies of CYR61 promoter/Enhancer3 were increased after transfection with GV219-mFOXA1 for 48 h in HCT116 cells, as assessed by 3C; significance determined by the paired-samples t-test. l Relative activity of promoter and enhancers after transfection of HCT116 cells with GV219-mFOXA1 for 48 h, as assessed by relative luciferase reporter gene activity. Significance for all data except a, b, f and k was determined by the independent samples t-test. Data are shown as mean ± S.D., n = 3. *P < 0.05, **P < 0.01, ***P < 0.001