Fig. 4

SIAH2 is a putative target for miR-146b. a Sequence alignment identified that the complementary region to the miR-146b seed region in the SIAH2 mRNA 3′UTR was conserved between human, mouse, rat, and chimpanzee; b miR-146b binds to and represses SIAH2 mRNA through the mRNA 3′UTR. pGL3-control luciferase vector containing the SIAH2 mRNA 3′UTR was co-transfected with indicated amounts of miR-146b mimic in HEK293 cells, and luciferase activities were analyzed 24 h post-transfection. A structurally unrelated miRNA (cel-miR-67) served as a negative control. c IEC-6 cells were treated with 50 ng/ml TGF-β with or without transfection with miR-146b inhibitor before treatment. After 48 h treatment, total SIAH2 protein and mRNA transcripts were analyzed by immunoblotting and real-time Q-PCR, respectively. Values are means ± SEM run in triplicates; letters indicate significant differences between treatment groups (P < 0.05)