Figure 4

LRP6 and PIK3R2 are direct targets of miR-126-3p. (A) MiR-126-3p and its putative binding sequence in the 3′UTR of LRP6 and PIK3R2. The mutant miR-126-3p binding site was generated in the complementary site for the seed region of miR-126-3p. (B) Hep-G2 and BEL-7402 cells were co-transfected with miR-126-3p mimics or NC and wild-type or its mutant-type (either LRP6 or PIK3R2), and the luciferase activities were examined. The firefly luciferase activity of each sample was normalized to the Renilla luciferase activity. (C) Western blot results of endogenous LRP6, PIK3R2, beta-catenin and p-AKT proteins in HCC cells transfected with miR-126-3p mimics or anti-mir-126-3p and its NC. (D) Analysis of LRP6 and PIK3R2 expression in subcutaneous tumors by immunohistochemistry. NC indicated negative control, **P < 0.01.