Fig. 5

FOXC1 promotes beta-catenin expression by enhancing the activity of beta-catenin promoter. a and b Real-time PCR and western blotting were used to detect changes in beta-catenin mRNA, GSK3β, pGSK3β and total and nuclear beta-catenin protein levels induced by FOXC1 knockdown or overexpression. c and d Representative immunohistochemical staining of FOXC1 and beta-catenin in xenograft tumors. e Luciferase reporter assays showed that FOXC1 significantly enhanced beta-catenin promoter activity. f Putative FOXC1 binding sites in the beta-catenin promoter and mutations in corresponding binding sites. Schematic representations of wild-type and mutant beta-catenin promoters (left) and the corresponding relative luciferase activity (right) are shown. g ChIP and real-time PCR assays were used to investigate the binding of FOXC1 to the putative binding site BS2. All experiments were independently repeated three times. The bar graph presents the mean ± SD. **P < 0.01