Figure 8.
ERαD258A prevents the nongenomic effect of E2 on phosphorylation of CSE and endothelial H2S release. A, HUVECs were transfected with empty vector (2 μg) or mutant ERα (D258A, 2 μg) plasmid for 48 h and treated with E2 for 15 min. p-VSAP, VASP, p-ERK, p-AKT, and ERα were detected by Western blotting. B, HUVECs were transfected with empty vector (2 μg) or mutant ERα (D258A, 2 μg) plasmid for 48 h and treated with E2 for 15 min. The interaction between ERα and Gαi was measured by immunoprecipitation and Western blotting. C and D, HUVECs were transfected with empty vector (2 μg) or mutant ERα (D258A, 2 μg) plasmid for 48 h and treated with E2 for 15 min, CSE phosphorylation level and total CSE was detected by immunoprecipitation and Western blotting. The medium H2S concentrations were measured (mean ± S.D.; n = 3 independent experiments; ***, p < 0.001 versus empty vector CON; ###, p < 0.001 versus empty vector E2, one-way ANOVA followed by the post hoc LSD test).