Figure 5.

Insig1 silencing inhibited FSH-mediated LHR upregulation. Rat granulosa cells were isolated from immature female rats injected subcutaneously with estradiol (1.5 mg) for 3 consecutive days. Cells were transfected with 50 nM of either negative CTLsi or siInsig1 and cultured for 48 hours. Cells were then treated with FSH (50 ng/mL) and E2 (1 nM) for 24 hours. Total RNAs were isolated and reverse transcribed, and the resulting complementary DNAs were subjected to real-time PCR quantitation using specific primers and probes for LHR. The graph represents changes in mRNA levels normalized to 18S ribosomal RNA and shown as fold change vs control. Error bars represent mean ± standard error. *P < 0.05 vs CTLsi; **P < 0.05 vs CTLsi + FE; n = 4.