Figure 4.
PKG-Iβ directly interacts with CSE and mediates E2-induced phosphorylation of CSE and endothelial H2S release. A and B, HUVECs were transfected with PKG-Iα (2 μg) or PKG-Iβ (2 μg) plasmid for 48 h, and treated with E2 for 15 min. The efficiency of overexpression was confirmed by Western blotting. C and D, CSE phosphorylation level and total CSE were detected by immunoprecipitation and Western blotting. E, HUVECs were transfected with PKG-Iα (2 μg) or PKG-Iβ (2 μg) plasmid for 48 h, and treated with E2 for 15 min. The medium H2S concentrations were measured (mean ± S.D.; n = 3 independent experiments; **, p < 0.01 versus empty vector CON, one-way ANOVA followed by the post hoc LSD test). F, the interaction between CSE protein and PKG-Iβ or PKG-Iα protein was detected by immunoprecipitation and Western blotting. Input refers to whole cell lysates, and IgG refers to normal rabbit IgG.