FIGURE 2.

The inhibition of iNOS induction mediated by Ang-(1-7) is dependent on Mas receptors. After exposing HASMC for 18 h to (A) Ang II (100 nM) or (B) IL-1β (2.5 ng/mL) with or without 100 nM Ang-(1-7), iNOS levels were determined by Western blot. In some experiments, cells were pre-incubated with the Mas receptor antagonist A779 (1 μM). (C) iNOS levels (red) were also determined by indirect immunofluorescence in response to the different stimuli. Nuclei were counterstained with DAPI (blue) (400×). In another set of experiments, nitric oxide (NO) production was determined in the supernatants of HASMC exposed for 18 h to Ang II (D) or IL-1β (E), with or without Ang-(1-7) and A779. The Mas receptor antagonist D-Pro⁷-Ang-(1-7) (1 μM) was also capable to block the inhibitory effect of Ang-(1-7) on iNOS induction by Ang II (F) or IL-1β (G). Results are expressed as mean ± SEM; n = 5. Representative blots are shown in the corresponding panels. ^∗p < 0.05 vs untreated cultures, ^#p < 0.05 vs Ang II- or IL-1β-treated cultures, respectively.