Fig. 4

CBP is recruited by FOXA1 to up-regulate CYR61 enhancer activity and CYR61 expression. a CBP enrichment decreased at Enhancer1 and Enhancer3 after treatment with FOXA1 siRNA for 72 h in RKO cells, and b increased at the promoter and Enhancer3 after FOXA1 over-expression for 48 h in HCT116 cells, as assessed by ChIP-qPCR. c CYR61 mRNA and eRNA, and d protein levels were decreased after treatment with CBP siRNA for 26 h in RKO and SW480 cell lines. e CBP and H3K27ac enrichment was decreased after treatment with CBP siRNA for 26 h in RKO cells, as assessed by ChIP-qPCR. f Relative crosslinking frequencies of CYR61 promoter/Enhancer1 were decreased after treatment with CBP siRNA for 26 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 with CBP siRNA for 26 h in RKO cells, as assessed by relative luciferase reporter gene expression. CBP and FOXA1 mRNA expression levels in RKO cells after treatment with CBP siRNA for 26 h h or with FOXA1 siRNA for 72 h i. CBP and FOXA1 protein expression in RKO cells after treatment with CBP siRNA for 26 h j or with FOXA1 siRNA for 72 h k. (L) FOXA1 enrichment after treatment with CBP siRNA for 26 h in RKO cells, as assessed by ChIP-qPCR. Significance for all data except f 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