Figure 5. LH/EGF promoted GPR30 protein accumulation in cumulus cells via a non-genomic pathway mediated by the activation of the EGF receptor. Mice were treated as shown in Figure 4, The proteins were detected using western blot (A), and the bands of GPR30 were quantified using gray scanning (B). PMSG-primed mice received EGF or EGF plus AG1478 (AG) for 8 h, proteins were detected by western blot (C) and the bands of GPR30 were quantified using gray scanning (D), the mRNA levels of GPR30 were measured using RT-qPCR (E). Immunostaining showed the fluorescence intensity (F) and the distribution (G) of GPR30 in the control, EGF, and EGF plus AG1478 groups, bar=50 μm. The amount of EGFR in COCs was detected by western blot (H and I), OOs indicate the oocytes and CCs indicate the cumulus cells. Data are presented as fold induction relative to the unstimulated control (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.