Fig. 4.

Effect of N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-(N,N-dimethylamino)acetamide hydrochloride (PJ34) on tissue poly(ADP-ribose) (PAR) content, respiratory complex subunits expression and mitochondrial DNA (mtDNA) content in Ndufs4 knockout (KO) mice. (A) The effects of a 10-day treatment (postnatal days 30–40) with PJ34 (daily intraperitoneal injections of 20 mg/kg) on tissue PAR content is shown. (B) Densitometric analysis of the effects of PJ34 on tissue PAR content of Ndufs4 KO mice. (C) mRNA levels of several mitochondrial [cyclooxygenase (COX)1, COX2, NADH dehydrogenase 2 (ND2)] and nuclear (NADH dehydrogenase (ubiquinone) flavoprotein 2 (NDUFV2), COX15, and ATP synthase, H+ transporting, mitochondrial F1 complex, delta subunit (ATP5D)) respiratory complex subunits in different organs of Ndufs4 heterozygous (HET) and KO mice. (D) The effects of PJ34 on transcripts levels of the respiratory complex subunits in KO mice are also shown. Succinate dehydrogenase complex, subunit A (SDHA) expression levels in different organs of (E) heterozygous and (F) KO mice treated or not with PJ34 is shown by Western blotting and (G) Densitometric analysis. (H) Effects of PJ34 on mitochondrial content (expressed as ND1/beta actin gene ratio) or (I) nicotinamide adenine dinucleotide (NAD) levels in different organs of Ndufs4 KO mice. Basal NAD content was 0.73 ± 0.12 μmol/g tissue, 0.64 ± 17 μmol/g tissue, 35 ± 0.08 μmol/g tissue, 0.1 ± 0.005 μmol/g tissue, 0.67 ± 0.21 μmol/g tissue, 0.59 ± 0.16 μmol/g tissue in the brain, pancreas, liver, spleen, heart, and skeletal muscle (sk. muscle), respectively. (A, E, F) A blot representative of 4 mice per group is shown. (B, C, D, G, H, I), columns represent the mean ± SEM of 4 mice per group. *p < 0.05, **p < 0.01, ***p < 0.001 vs vehicle, analysis of variance plus Tukey’s post hoc test