Figure 3

CADM1 was a direct target of miR-1246. (A) Putative binding site for miR-1246 in the 3′UTR of CADM1 was revealed by TargetScan. (B) The miR-1246 binding site on CADM1 3′UTR was confirmed by luciferase assay in 293 T cells after cotransfection with (i) a plasmid containing a fragment of CADM1 3′UTR that included either the wild type or mutant predicted miR-1246 binding site and (ii) the miR-1246 mimic or the mimic control. Data represent the mean ± SD of at least three independent experiments. *P < 0.01. (C) Western blot assay showed increased CADM1 expression in SMMC7721 cells after transfection with the miR-1246 inhibitor (800 nM). (D) Real time PCR showed that miR-1246 did not affect CADM1 mRNA expression.