EGF downregulates the expression but causes protein accumulation of GPR30 in COCs cultured in vitro. COCs isolated from PMSG-primed mice were treated with or without EGF for 0, 2, 4, 8, and 16 h. After that, GPR30 mRNA levels were measured using RT-qPCR (A). The amount of GPR30 protein in COCs cultured with EGF was detected using western blot (B), and the bands were quantified using gray scanning (C). COCs were cultured in vitro with or without EGF for 8 and 16 h, GPR30 protein levels were detected using western blots (D), and the bands were quantified using gray scanning (E). Data are represented as fold induction relative to the unstimulated control (0 h) in panels (A), (C) and (E), and bars are presented as average±SEM. In panels A and E, “*” indicated that p<0.05, and “**” indicated that p<0.01. In panel C, different lowercase letters indicate significant differences between groups (p<0.05). Three independent replicates were performed for each experiment.