Research Paper Volume 12, Issue 20 pp 20801—20816

EGFR positively regulates GPR30 protein stability. Primary FGCs were treated with EGF to determine whether GPR30 accumulated after EGFR activity. (A) Western blot of proteins in FGCs after EGF treatment. (B) Graphical representation of the quantification of GPR30 protein levels shown in panel A. (C) Immunolabeling of the FGCs with GPR30 antibody (green) and DAPI (blue), bar=10 μm. (D) Western blot of GPR30 in FGCs treated with CHX only. (E) Western blot of GPR30 in FGCs treated with CHX plus EGF. (F) Graphical representation of the quantification of GPR30 protein levels shown in panels D (con) and E (EGF) to determine protein half-life. Data are presented as fold induction relative to the unstimulated control (0 h). The bars of panels B and F are presented as average±SEM. Different lowercase letters indicate significant differences between groups (p<0.05). Three independent replicates were performed for each experiment.