Fig. 6: Activation of Wnt/β-catenin signaling induces EIF3H expression.

a qRT-PCR analysis of EIF3H mRNA in DLD1 and HCT116 cells treated with L-Wnt3a-expressing cell conditioned medium. Data are presented as the means ± SD, n  =  3 biologically independent experiments. b qRT-PCR and western-blot analysis of EIF3H and AXIN2 levels in DLD1 and HCT16 cells treated with different concentrations of Wnt pathway inhibitor ICG-001. Data are presented as the means ± SD, n  =  3 biologically independent experiments. c qRT-PCR and western-blot analysis of EIF3H and AXIN2 levels in DLD1, HCT16, and RKO cells treated with different concentrations of Wnt pathway inhibitor NCB0846. Data are presented as the means ± SD, n  =  3 biologically independent experiments. d, e qRT-PCR d and western-blot e analysis of EIF3H and c-Myc level in DLD1 and HCT116 cells cultured with L-Wnt3a-expressing cell CM, in the presence or absence of Wnt pathway inhibitor NCB0846 (10 µM). Data are presented as the means ± SD, n  =  3 biologically independent experiments. f The potential binding sequence of TCF4 obtained from JASPAR website. g Deletion mutants of the EIF3H promoter (middle); HCT116 cells were subjected to chromatin immunoprecipitation using antibodies against IgG, TCF4, or β-catenin, followed by qRT-PCR for the loci on EIF3H promoter (right). NLIF3 was a positive control. Data are presented as the means ± SD, n  =  3 biologically independent experiments. h, i CRC line DLD1, HCT116 and RKO cells were transfected with Flag-TCF4 or Flag-β-catenin plasmids. qRT-PCR and western-blot analysis of EIF3H level. Data are presented as the means ± SD, n  =  3 biologically independent experiments. j DLD1 and HCT116 cells were treated with different concentrations of Wnt pathway inhibitor LGK974 plus MEK1/2 inhibitor trametinib. The colony formation assay was performed. The p values were determined by unpaired two-tailed t test for a–d, g, h. Representative immunoblots shown in the figures were repeated three times independently with similar results. Source data are provided as a Source Data file.