Mice were subjected to Intraperitoneal injection of BCNU (20 mg/kg/day) for 4 days. Fractions of mitochondria and cytosol were isolated from the hearts. A, Immunoblotting analysis of mitochondria using anti-SOD2 (upper panel, titer used: 1 mg/ml, 4,000X) and anti-GR (lower panel, titer used: 1 mg/ml, 1,000X) polyclonal antibodies. B, Analysis of GR activities from the cytosolic and mitochondrial fractions. C, State 3, state 4, and FCCP-mediated OCRs of isolated mitochondria from SOD2-tg and wild type mice. D, Respiratory control index obtained from the ratio of state 3 and state 4 OCRs. E,
a and b: •O2− generation mediated by mitochondria isolated from murine hearts of BCNU-treated wild type and SOD2-tg mice; c and d: the same as a and b, except that antimycin A (10 μM) was included in the system. *p<0.05, assessed by student’s t-test between WT control and BCNU-treated WT or SOD2-tg control and BCNU-treated SOD2-tg in B, C, and D. Comparison among four groups (WT, BCNU-treated WT, SOD2-tg, and BCNU-treated SOD2-tg) in D was analyzed by one-way ANOVA followed by Tukey’s post-hoc test, indicating there is no significant difference among the four groups (p = 0.16222).