Fig. 5

Validation miR-92b-3p directly binds to PWRN2 and the direct interaction between miR-92b-3p and its predictive target gene (TMEM120B) by the dual luciferase activity assay. a Interaction regions of PWRN2/miR-92b-3p and miR-92b-3p/TMEM120B predicted by miRanda software. MiR-92b-3p, which is predicted to process the MREs in PWRN2, also has a binding site in the 3′-UTR of TMEM120B. b The relative luciferase activity was assayed following co-transfection of miR-92b-3p mimics with the constructs encoding the wild-type or mutant PWRN2 cDNA into the cumulus cells. c The relative luciferase activity was assayed following co-transfection of miR-92b-3p mimics with the constructs encoding the wild-type or mutant miR-92b-3p binding site of TMEM120B 3′-UTR into the cumulus cells. The constructed reporter vectors (WT or MUT) are shown on the x-axis, and the y-axis represents the normalised luciferase activity (firefly luciferase activities were normalised to renilla luciferase activities). The experiments were performed independently in triplicate. * indicates P < 0.05, ** indicates P < 0.01, NS indicates no significant difference. The results are presented as means ±S.E.M