Figure 6.

FeTMPyP (2.5 µmol/L) has no significant effect on nitrotyrosination of mitochondrial proteins but decreases peroxynitrite (PN) production. A: representative Western blot for nitrotyrosine content and VDAC (loading control). B: representative bar diagram. C: mitochondrial PN production. Mitochondria were isolated from wild-type WT and endothelial NO synthase knockout (eNOS-KO) mice (n = 4 mice for Western, n = 4–8 mice for PN measurements) brains, treated with FeTMPyP (2.5 µmol/L) for 60 min. Mitochondrial pellets were solubilized with NP 40 buffer and subjected to SDS-PAGE (4%–20% gradient). Nitrotyrosine content in the proteins was detected on anti-nitrotyrosine Immun-Blot using enhanced chemiluminescence. Total band intensity was measured using ImageJ. PN measurements were taken using DAX-J2 PON green (excitation: 490 nm and emission: 530 nm) Data are presented as means ± SE and analyzed by repeated-measures two-way ANOVA. *P < 0.05, statistically significant. C, Control; S, FeTMPyP.