FIGURE 7.

NoxA1ds disrupts VEGF-stimulated Nox1-NOXA1 interaction. FRET between Nox1-YFP and NoxA1-CFP transfected HPAEC in the presence or absence of 20 nm VEGF and/or 10 μm NoxA1ds. Relative fluorescence of CFP is green, and YFP is red. Traces below the images indicate fluorescent intensities of CFP and YFP below the arrow overlaid on each cell. A, transfected HPAEC were treated with vehicle for 1 h prior to imaging cells; photobleaching of Nox1-YFP was complete and resulted in a concomitant increase in CFP fluorescence. B, transfected HPAEC were treated with 20 nm VEGF for 1 h prior to imaging cells; photobleaching of Nox1-YFP was complete and resulted in an increase in CFP fluorescence of greater magnitude than without VEGF. C, transfected HPAEC were treated with 10 μm NoxA1ds peptide for 1 h prior 20 nm VEGF for 1 h. Cells were then imaged after VEGF treatment. Photobleaching of Nox1-YFP was complete but did not result in a concomitant increase in CFP fluorescence. D, quantification of FRET efficiency from images A–C. Values expressed as n = 6, two separate experiments; *, p < 0.05 versus vehicle; ***, p < 0.001 versus VEGF, one-way ANOVA and Bonferroni post-test.