Fig. 3

The effect of LH on AREG expression in human theca cells and the role of the cAMP/PKA signalling pathway in the process. A LH stimulated both the gene expression of AREG in theca cells (a1) and the secretion of AREG in theca cell culture medium (a2) in a dose-dependent manner. Cells were cultured for 6 h with or without different concentrations of LH. B Time course of the expression of AREG in the presence of LH (400 mIU/mL) in human theca cells. The mRNA levels of AREG were measured by quantitative PCR (b1); the levels of AREG protein in the culture medium were measured by ELISA (b2). C, D, and E Relative expression of AREG, EGFR, and LHCGR in human theca cells treated with LH (400 mIU/mL, with or without 10µM H89), forskolin (10 µM), or db-cAMP (500 µM) for 6 h. The mRNA levels of AREG were measured by quantitative PCR; the levels of AREG protein in the culture medium were measured by ELISA (C and D); the levels of AREG protein in theca cells were measured by western blot (D). In A and B, each experiment was performed once. In C-E, each experiment was repeated three times. *P < 0.05, ** P < 0.01, ***P < 0.001