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Figure 5 Enhancement of endothelial-type nitric oxide synthase (eNOS) promoter activity in EA.hy 926 cells after incubation with French red wines. (A) EA.hy 926 cells were stably transfected with p-eNOS-3500-Hu-Luc-neo. Stable cells were incubated for 24 h with ethanol (EtOH-Co, 1.25%, v/v), or various concentrations of two French red wines ("L.Chevaliers" and "Ch.Bonnet reserve"). The luciferase activity (normalized by protein content) was taken as a measure of eNOS promoter activity. Bars represent means ± SEM of three independent experiments. Asterisks indicate significant differences from ethanol-treated cells (**p < 0.01; ***p < 0.001; by ANOVA followed by the Fisher PLSD test). (B) EA.hy 926 cells were transiently transfected with pGl₃-Basic (containing a promoterless luciferase gene) or different eNOS promoter luciferase constructs (p-eNOS-1600-Hu-Luc, p-eNOS-1111-Hu-Luc, p-eNOS-633-Hu-Luc, and p-eNOS-326-Hu-Luc, containing 1.6-kb to 0.33-kb of the eNOS promoter cloned before a luciferase reporter gene). The different transfected cells were either exposed 24 h to ethanol (EtOH-Co, 1.25%, v/v) or to the red wine "L.Chevaliers" (10%, v/v). The relative luciferase activity (corrected with renilla-luciferase activity; see Methods) was taken as a measure of eNOS promoter activity. Data represent means ± SEM of three independent experiments.