Fig. 5

SMAD4 served as a target of miR-486-5p. a The fragments of SMAD4 3′UTR-WT and SMAD4 3′UTR-MUT were presented. b Dual-luciferase reporter assay indicated that the luciferase activity of SMAD4 3′UTR-WT could be reduced by miR-486-5p (P < 0.0001). c RIP assay suggested that the enrichment of miR-486-5p and SMAD4 was increased in Ago2 (P < 0.0001). d Biotin-labeled RNA pull-down assay showed that the enrichment of SMAD4 could be pulled down by the bio-miR-486-5p probe (P < 0.001). e, f QRT-PCR and WB analysis suggested that the mRNA and protein levels of SMAD4 were promoted in posterior capsular tissues from PCO patients compared to normal humans. g Pearson correlation analysis revealed that miR-486-5p expression was negatively correlated with and SMAD4 expression in posterior capsular tissues of PCO patients. h, i QRT-PCR and WB analysis showed that the mRNA and protein levels of SMAD4 were increased in SRA01/04 cells treated with TGF-β2 in a dose-dependent manner (P < 0.05, P < 0.01, P < 0.0001). j QRT-PCR analysis revealed that the expression of miR-486-5p was enhanced by miR-486-5p mimic (P < 0.0001) while reduced by miR-486-5p inhibitor (P < 0.001) in TGF-β2-stimulated SRA01/04 cells. k WB analysis presented that the protein level of SMAD4 was decreased by miR-486-5p overexpression (P < 0.0001) and increased by measured by miR-486-5p inhibition (P < 0.0001). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001