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. 2020 Apr 20;21(8):2868. doi: 10.3390/ijms21082868

Figure 2.

Figure 2

GRK2 but not GRK5 desensitizes G protein-coupled estrogen receptor (GPER) in cardiac myocytes. (A) Immunoblotting for GRK2 and GRK5 in GPER immunoprecipitates from H9c2 cardiomyocyte extracts prepared after 15-min-long treatments with vehicle (DMSO, Veh), 100 nM G1, or 100 nM aldosterone (Ald). Blots for GPER are also shown to confirm equal amounts of receptor immunoprecipitated. IP: immunoprecipitation; IB: Immunoblotting; IgG: Negative control for the co-IP (general rabbit IgG was used in the IP instead of an anti-GPER antibody). (B) Total GTPγS binding to measure G1-induced GPER desensitization in H9c2 cardioymyocytes: Cells were treated with 100 nM G1 for 15 min following a 30-min-long pretreatment with either 100 nM G1 alone (G1-pretreated) or 100 nM G1 in the presence of 30 µM Cmpd101 (Cmpd101 + G1-pretreated). *, p < 0.05; n = 5 independent experiments performed in triplicate.