Fig. 2. IDH2 deficiency exacerbates hydrogen peroxide formation and oxidation of DNA in the kidney after cisplatin administration.
Idh2‒/‒ mice and wild-type (Idh2+/+) littermates were intraperitoneally injected with either cisplatin (C, 20 mg/kg B.W.) or 0.9% saline (vehicle, V) once. Some mice were treated with Mito-T (M, 0.7 mg/kg B.W.) daily, beginning 7 days before cisplatin injection and continuing until experiments were completed. Kidneys were harvested 3 days after cisplatin injection. a H2O2 was measured in whole kidney tissue (n = 4-5). b Kidney sections were immunostained with an anti-8-hydroxy-2′-deoxyguanosine (8-OHdG) antibody. c 8-OHdG-positive area (%) was measured. More than 10 fields per kidney section were analyzed (n = 3). d Fractions were confirmed by western blot analysis using anti-MnSOD for the mitochondria, -CuZnSOD for the cytosol, and -histone H1 for the nucleus. e, h Expression levels of Prx-SO3 and 4-hydroxynonenal (4-HNE) were determined in the mitochondrial fraction of kidneys by western blot analysis. f, h Band densities were normalized to COX IV band densities using the ImageJ program. COX IV bands in “e” and “g” are same band produced by same blot. i Expression of 4-HNE was detected in cytosolic fraction of kidneys by western blot analysis. GADPH was used as a loading control. j Band density was normalized to GAPDH band density using the ImageJ program. Results are expressed as means ± SE (n = 3–5 per group). Scale bars: b 50 µm. *p < 0.05 vs. respective V; #p < 0.05 vs. respective C; §p < 0.05 vs. V in Idh2+/+; †p < 0.05 vs. C in Idh2+/+. Nu nucleus, Mito mitochondria, Cyto cytosol