Fig. 3

miR-30a-5p and miR-30a-3p inhibit ZEB2 and mediate the p53 control over ZEB2. a Alignment of ZEB2 3’UTR with miR-30a-5p and miR-30a-3p (Targetscan). Nucleotides mutagenized to disrupt the miRNA/mRNA base pairing are underlined. b Immunoblots showing the modulation of ZEB2 in response to miR-30a (5p, 3p or both) ectopic expression and c after anti-miR-mediated inhibition of miR-30a. ZEB2 relative levels, normalized over Tubulin (loading control), are reported below. d miR-30a targets the 3′UTR of ZEB2: luciferase activity of the ZEB2 3’UTR reporter, wild-type (WT) or mutated in the miR-30a-5p and -3p binding sites (5pMUT and 3pMUT), was measured 48 h post transfection of MDA231 cells with the indicated miRNAs. miRCTR was set as a reference. CMV-Renilla was used for normalization. Results represent the mean value ± SD of three experiments. The asterisks (*) indicate the comparisons of miR-30a vs miRCTR that are statistically significant (p < 0.05). e miR-30a-5p and miR-30a-3p expression in MDA157 stably transduced with p53 (pLenti-p53+) or pLenti-GFP (pLenti-p53−), in the absence (anti-miR-30a−) or presence (anti-miR-30a+) of an anti-miRNA targeting miR-30a. Immunoblots for ZEB2, p53 and Vinculin (loading control) are shown on the right. Numbers below the blot indicate ZEB2 relative levels normalized over Vinculin