FIGURE 6.

Effects of thyrostimulin on cAMP, c-fos gene, and progesterone biosyntheses in granulosa cells treated with or without gonadotropins. Granulosa cells from immature rats (left panels) or from rats injected with PMSG for 12 h (middle panels) or with PMSG for 48 h followed by hCG for 12 h (right panels) were treated with thyrostimulin (indicated as A2/B5) in the presence or absence of gonadotropins as indicated. For the cAMP (A–C) and progesterone (G–I) assays, cells were incubated for an additional 48 h before measurements. For c-fos gene induction (D–F), cells were serum-starved for 20 h before thyrostimulin and gonadotropin treatments for an additional 30 min. Cells were then collected for cDNA preparation. The c-fos gene levels were quantified by real-time PCR and normalized against the β-actin expression level. At least three individual repeats were done. Data were obtained from triplicate experiments and are shown as the means ± S.D. *, p < 0.05.