Fig. 4

TCDD and DIM suppress the pro-metastatic factor SOX4 by inducing the miR-212/132 cluster. a 10 nmol/L TCDD and 25 μmol/L DIM suppressed SOX4 gene expression as quantified by real-time PCR and western blot. b Knockdown of Ahr by siAhr blocked the inhibitory effects of TCDD and DIM on SOX4 compared with siNS-transfected MDA-MB-231 and T47D cells. c and d Co-transfection of 3′UTR-SOX4-luc and miRNA antisense, as-miR-132 or as-miR-212, mitigated the inhibitory effects of TCDD and DIM on the luciferase activity in MDA-MB-231 and T47D. e SOX4 is a potential target gene of the miR-212/132 cluster; complementary binding site of miR-212/132 on the SOX4 3′UTR. f Co-transfection of 3′UTR-SOX4-luc and miRNA mimics, miR-132 or miR-212, suppressed the luciferase activity in MDA-MB-231 and T47D. g MiRNA mimics did not decrease the luciferase activity when co-transfected with 3′UTR-SOX4-luc that contains mutated binding sites for miR-212/132. Data are shown as mean ± SD of three independent experiments performed in triplicates. a *P < 0.05, significantly different from DMSO-treated control. b–d *P < 0.05, significantly different from the corresponding DMSO- and siNS-treated control or Ahr agonist- and miRNA antisense treatments. f and g *P < 0.05, significantly different from siNS-transfected control