Fig. 5.

MiR-200b-3p disrupts stability of c-Myc protein by inhibiting AKT2/GSK3β pathway. a Detection of c-Myc protein by western blot and miR-200b-3p by qPCR after overexpression of c-Myc and co-expression of c-Myc and miR-200b-3p in Caco2 cells. The grey value of c-Myc was normalized to that of the corresponding GAPDH (***p < 0.001). b Total GSK3β, p-GSK3β (Ser9), total c-Myc, p-c-Myc (S62) and p-c-Myc (T58) protein levels were detected by western blot in LoVo/miR, SW480/Zip-miR and SW480/Zip-miR cells with Tws119 treatment for 72 h. c AKT (1/2), AKT1 and AKT2 were detected by western blot in LoVo/miR and SW480/Zip-miR cells. d Predictive binding sites and mutant sites of miR-200b-3p to 3′UTR of AKT2 mRNA. e The luciferase activities of wild-type and mutant-type pmirGLO-3′UTRs of AKT2 mRNA in 293T cells after transfection of miR-200b-3p mimics (***p < 0.001). f AKT2, p-AKT2 (Ser474), total GSK3β, p-GSK3β (Ser9), total c-Myc, p-c-Myc (S62) and p-c-Myc (T58) protein were detected by western blot after transfection of miR-200b-3p mimics