Fig. 5: Complex I inhibition, in vitro and in vivo, is associated with an inflammatory gene response caused by increased oxidative stress.
From: Defective NADPH production in mitochondrial disease complex I causes inflammation and cell death
Immunoblots showing oxidative stress-mediated activation of ASK1/P38/JNK axis specifically in ND1 mutant cells a cultured in galactose or b after PPP inhibition using 6-AN at 100 μM for 48 h. Pro-inflammatory gene expression signature is induced in c 48 h galactose-grown or d PPP-inhibited ND1 cells and rescue by ME1 overexpression (n = 3). e Metabolomic analysis in brain samples of WT and Ndufs4 KO mice. Note that levels of GSH are reduced, while no changes are observed in ATP. GSH/GSSG ratio for WT is 4.3920 +/− 0.9039 (Average +/− Standard) (n = 5). f Increased phosphorylation of JNK (including long and short exposures) and g induced gene expression of inflammatory markers in the brain of Ndufs4 KO mice. h Reduced formate production using brain-isolated mitochondria from WT or Ndufs4 KO mice (n = 4). Immunoblots shown are representative of >3 independent experiments, and all other experiments are represented as means ± SEM., n > 3 biological replicates. Asterisks denote *p < 0.05, **p < 0.01, or ***p < 0.001. Two-way ANOVA in c, d, h. Paired two-tailed Student’s t test in e, g and one-way ANOVA in h. gluc/g glucose, Galac/G galactose, Pir Piericidin.
