Figure 7

JIB-04 inhibited the β-catenin signaling pathway in colorectal cancer cells. (A) mRNA expression levels of β-catenin target genes associated with CSCs after treatment with DMSO, 2 µM JIB-04, or 10 µM salinomycin for 24 h. The mRNA levels of the target genes were normalized to that of GAPDH. The expression level of DMSO-treated cells was set as 1 (n = 3). (B) Protein expression of LGR5 and CD44 after treatment with DMSO or 2 µM JIB-04 for 24 h. GAPDH was used as a loading control. (C) TOP flash reporter assay of β-catenin-based promoter activity after treatment with DMSO, 2 µM JIB-04, or 10 µM salinomycin for 24 h. Cells were transfected with TOP or FOP flash plasmids and renilla luciferase plasmid. Luciferase activity was measured 48 h after transfection and normalized to that of renilla. The luciferase activity of DMSO-treated cells was set as 1 (n = 3). (D) Western blot analysis of histone methylation levels after treatment with DMSO or 2 µM JIB-04 for 24 h. H3 was used as a loading control. *p < 0.05, **p < 0.01, ***p < 0.001, compared with DMSO.