Functional characterizations of two IGF2BP3-regulated targets in CRC. (A, B) The mRNA levels of MAP2K1 (A) and TPR (B) were analyzed in two independent CRC patient cohorts. (C) HCT116 cells were transfected with siRNA targeting MAP2K1 and a nontargeting siRNA control, and the knockdown efficiency of MAP2K1 was confirmed by western blotting analysis (left panel). The effect of MAP2K1 silencing on the proliferation of HCT116 cells was analyzed with a CCK-8 assay (right panel). (D) HCT116 cells were transfected with siRNA targeting TPR and a nontargeting siRNA control, and the knockdown efficiency of TPR was confirmed by western blotting analysis (left panel). The effect of TPR silencing on the proliferation of HCT116 cells was analyzed with a CCK-8 assay (right panel). (E, F) The SW480 cells overexpressing IGF2BP3 were further transfected with siRNA targeting MAP2K1 (E) and TPR (F). The expression of IGF2BP3, MAP2K1 and TPR was analyzed with RT-qPCR analysis (E, F left panel). The proliferation of the indicated cells was examined with a CCK-8 assay (E, F right panel). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.