Granulosa cells were isolated from immature female rats injected subcutaneously with estradiol (1.5 mg) for three consecutive days. Cells were cultured in serum-free media for 24 hours and then treated with FSH (50 ng/ml) and E2 (1nM) for 2, 4, 6, 12, 24, and 48 hours. Total RNAs were isolated and reverse transcribed, and the resulting cDNAs were subjected to real-time PCR quantitation using specific primers and probes for LHCGR (A) or miR-122 (B). The graph represents changes in mRNA levels normalized to 18S rRNA or U6 SnRNA and shown as fold-change vs. control. Error bars represent mean ±SE. *p<0.05 vs. control; n=4.