Fig 2. Mitochondrial DNA integrity and expression of mtDNA-encoded genes in chagasic mice (± PARP1).
Mice (WT and PARP1-/-) were infected with T. cruzi and sacrificed at 150 days’ pi. (A) Representative gel images (n = 3 mice/group) show myocardial levels of 10 kb mtDNA with short 177-bp mtDNA and 96-bp GAPDH (nuDNA) fragments as controls. The PCR amplification was conducted for 28 cycles. (B) Densitometry analysis was performed on PCR gels representing n? 6 mice/group, and density of the 10 kb mtDNA band was normalized against mtDNA (B.a) and nuDNA (B.b) fragments. (C-G) Myocardial mRNA levels for mtDNA-encoded genes were determined by RT-qPCR, and normalized to GAPDH mRNA (n ? 5, triplicate observations per mouse). (H-K) Representative Western blotting images (n = 3 mice per group) of cardiac mitochondrial levels of COI and AMPK-like (COIV, loading control) and total heart homogenate levels of AMPK (GAPDH, loading control) are shown (H). Densitometry analysis was performed on gel images for n = 6 mice for each protein, and data for COI and AMPK-like levels in mitochondrial fractions (I&J) and AMPK levels in total heart homogenates (K) were normalized to COIV and GAPDH, respectively. In bar graphs, data are plotted as mean value ± SEM. Statistical significance are marked as *WT.Tc vs. WT, &PARP1-/-.Tc vs. PARP1-/-, and # WT.Tc vs. PARP1-/-.Tc (*,#p<0.05, ***,&&&,###p<0.001).