Figure 7. Effect of regulator of G‐protein signaling 5 deficiency (RGS5) on endothelial NO synthase (eNOS) expression in human brain microvascular endothelial cells (HBMECs) and brain tissue before and after middle cerebral artery occlusion (MCAO).

A, Western blotting analysis and quantification of eNOS phosphorylation and expression in control and RGS5 knockdown (RGS5‐KD) HBMECs, with and without oxygen glucose deprivation and l‐glutamate treatment (OGD+l‐glutamate; 1 mmol/L for 3 hours) (n=5). NO production measured in whole brains before MCAO (B) and in contralateral and ipsilateral hemispheres after MCAO (C) of controls and endothelial Rgs5 deficient (EC‐Rgs5^–/–) mice (n=5). Rho‐associated kinase (ROCK) activity, measured by myosin phosphatase target subunit 1 (MBS) phosphorylation (D) and eNOS phosphorylation (E) and expression in contralateral and ipsilateral hemisphere after MCAO of control and EC‐Rgs5 ^–/– mice. F, DAF‐FM staining of EC‐Rgs5^–/– and control mice coronal brain sections 24 hours after MCAO (n=3). Bar=1 mm. For all data sets, 1‐way ANOVA (B) or 2‐way ANOVA (C through E; variables, ipsilateral vs contralateral and type of mice) followed by post hoc Tukey tests. Nonnormally distributed data sets were analyzed by Kruskal‐Wallis tests (A). *P<0.05, **P<0.01, #P<0.001. Control indicates EC‐Rgs5^flox/flox without tamoxifen; Control+TMX, Cdh5‐Cre^ERT2 with tamoxifen; and EC‐Rgs5^–/–, endothelial cell–specific RGS5‐deficient mice.