Figure 9. EGFR protein was increased by GPR30 activation in COCs.PMSG-primed female mice were injected with hCG, and the COCs in the ovaries were collected at 0, 2, 4, 8, and 16 h. After that, the EGFR protein levels were evaluated using western blot (A) and quantified using gray scanning (B). COCs were cultured in vitro with EGF treatment for 0, 2, 4, 8, and 16 h, EGFR protein was detected using western blot (C) and quantified using gray scanning (D). In the next experiment, 17β-E2, G1, G15 and ICI182780 were used to explore the mechanism of estrogen in the maintenance or even upregulation of EGFR levels in COCs cultured in vitro for 8 h. EGFR protein levels were detected using western blot (E) and quantified using gray scanning (F). Data are represented as fold induction relative to the unstimulated control (0 h or con). Bars are presented as average±SEM. Different lowercase letters indicate significant differences between groups (p<0.05). Three independent replicates were performed for each experiment.