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. Author manuscript; available in PMC: 2009 Jan 9.
Published in final edited form as: Circ Res. 2008 Jul 3;103(4):360–368. doi: 10.1161/CIRCRESAHA.108.178897

Figure 5.

Figure 5

Rac1 mediates the expression of eNOS mRNA through transcriptional and posttranscriptional mechanisms. A, Effect of Rac1 inhibition on eNOS mRNA and protein levels in cultured ECs as assessed by Northern and immunoblot analyses. Left panel, human aortic endothelial cells (HAEC) were infected with adenoviruses encoding enhanced green fluorescent protein or dominant negative Rac1 (N17Rac1) at 50 MOI for 24 hours. Right panel, bEND.3 cells were treated without or with 100 μmol/L Rac1 inhibitor (NSC23766) for 24 to 72 hours. 18S: EtBr-stained bands corresponding to 18S rRNA. B, Effect of Rac1 inhibition on eNOS promoter activity. Left panel, the structure of luciferase reporter construct harboring -1.6 kb upstream sequence of human eNOS gene (F1. Luc). Right panel, the promoter activity of F1. Luc in BAEC and bEND.3 cells cotransfected with pcDNA3, N17Rac1, or dominant negative PAK1 (K299R PAK1) expression vector and subsequently treated with vehicle, Rac1 inhibitor, apocynin (30 μmol/L) or NAC (10 mmol/L). Data are mean±SEM of the relative luciferase activity from triplicate assays. C, Effect of Rac1 inhibition on eNOS mRNA stability. Confluent bEND.3 cells were treated with 5,6-dichlorobenzimidazole 1-β-d-ribofuranoside (50 μmol/L) in the absence or presence of 100 μmol/L Rac1 inhibitor for indicated periods. eNOS mRNA expression was assessed by Northern blot analysis (left) in duplicate and quantified by densitometry (right panel). *P<0.05; #P<0.01.