Fig. 3

The expression of FoxM1 is regulated by Hh-Gli1 signaling. a Western blot analysis of the Gli1, FoxM1, and CCNB1 protein levels in six CRC cell lines. b Real-time PCR analysis of the Gli1, FoxM1, and CCNB1 mRNA expression levels in six CRC cell lines. The HT29 cells’ mRNA expression level was used as the normalized control. c Detection of Gli1, FoxM1, and CCNB1 protein expression in Caco2 cells after treatment with Hh signaling pathway activator. Caco2 cells were treated with 1 μM or 2 μM purmorphamine for 48 h, lysed and subjected to a Western blot analysis. d-f The inhibition of the Hh signaling pathway inhibited the protein expression of Gli1, FoxM1, and CCNB1, as demonstrated by the Western blot. HCT116 cells were transfected with Gli1-miRNAi or control miRNAi constructs for 48 h (d) or treated with the Gli inhibitor GANT61 (e) or Smo inhibitor cyclopamine (f) at the indicated time and concentrations. The cells were lysed and subjected to Western blotting. g Real-time PCR analysis of Gli1, FoxM1, and CCNB1 mRNA expression in HCT116 cells after treatment with GANT61 or cyclopamine. The mRNA expression levels were normalized to that of β-actin and expressed as fold change compared with the DMSO control. Error bars represent the mean and S.D. of three independent experiments. miR-Gli1: miRNAi-Gli1. *, p < 0.05; **, p < 0.01