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)..
“Some people who became sick did not report eating romaine lettuce, but had close contact with someone else who got sick from eating romaine lettuce.”
“It often includes investigating the multiple steps along the way. These steps can include suppliers, distributors and processors where the lettuce was chopped and bagged, and then back to the farm or farms that could have grown the lettuce that ended up in those bags. It’s a labor-intensive task.”