(A) HCT116 cells were treated with Wnt3a for various times and then harvested for qRT-PCR to assess mRNA expression of TGIF1, CCND1 and AXIN2. (B) After LoVo cells were transfected with shControl and β-catenin shRNA for 24 hours, mRNA expression of β-catenin, TGIF1 CCND1, c-Myc and AXIN2 were examined by qRT-PCR. *, p <0.05; **, p <0.01; ***, p<0.001.
(C) Protein expression of β-catenin, TGIF1 and c-Myc in CRC tissues (Tumor) and paired normal tissues (Normal) were tested by Immunohistochemistry. Scale bars=100μm. (D) Expression correlation between TGIF1 and β-catenin or TGIF1 and c-Myc in CRC samples. (E) Immunohistochemistry was performed in the xenografts derived from control and TGIF1 knockdown LoVo cells. Scale bars=100 μm. (F) Expression of Wnt target genes (CCND1, AXIN2 and c-Myc) was performed in the xenografts derived from control and TGIF1 knockdown LoVo cells. Error bars indicate S.E. of the values of each group. *, p<0.05; **, p<0.01.