Fig. 6

Knockdown of Nurr1 partially attenuates the effects of miR-132, and re-expression of Nurr1 abrogates the stimulatory effect of miR-132 on E2 synthesis. a mGCs were transfected with 100 nM siRNA targeting Nurr1 or negative control siRNA. Real-time PCR detection of Nurr1 mRNA levels was performed 48 h after transfection of mGCs. b After knockdown, Nurr1 protein levels were analyzed by Western blot. c siRNA was transfected 24 h before transfection of miR-132 mimics or negative controls, followed by continuous culture for an additional 48 h. The culture medium was collected for the measurement of E₂ levels 24 h and 48 h after transfection. d Total RNA was extracted from mGCs and Cyp19a1 mRNA levels were measured using real-time PCR. e Culture medium was collected for the measurement of E₂ levels 48 h after transfecting mGCs with the Flag-Nurr1 or the Flag-empty vector (EV) plasmids and miRNA mimics, as indicated. The results represent the mean +/-SEM of three independent experiments performed in triplicate. Values with different superscripts (a, b, c) are significantly different (p < 0.05). *p < 0.05; **p < 0.01; N.S. no significant difference, compared with the negative control (NC)