Figure 4.

Perm1 enhances mitochondrial biogenesis in neonatal mouse cardiomyocytes. Primary cultured neonatal mouse cardiomyocytes (NCM) were infected with adenoviruses expressing LacZ or Perm1. Forty-eight hours later, the cells were harvested. A and B, whole cell protein lysates were subjected to western blot using antibodies as indicated. OxPhos protein levels were quantified with expression level of LacZ (control) infected cells = 1. C, the relative mtDNA content was determined in Perm1-infected cells, relative to that detected in control (LacZ)-infected NCM. (LacZ expression was set =1.) D and E, mRNA levels for the indicated mitochondrial genome-encoded OxPhos genes mt-CoxII and mt-CoxIII and other OxPhos component genes, NDUFS3, SDHb, UQCRC2, COXIV, and ATP5b were determined by RT-qPCR, normalized to 36B4 levels, and expressed relative to levels of each gene in control (LacZ) NCM, set = 1. B–E, data are the mean ± SD, expressed relative to LacZ. ∗p < 0.05 versus LacZ. Data are the mean of six experimental replicates from two representative experiments. F, oxygen consumption rates of NCM were measured in the absence and presence of 1 μM oligomycin (Oligo), 800 nM FCCP, and 1 μM rotenone/antimycin (RAA). Rates are normalized by 20 × 10³ cells. Data are the mean ± SD. ∗p < 0.05 versus LacZ. Data are the mean of nine experimental replicates from two representative experiments. Data are the mean ± SD, expressed relative to LacZ. ∗p < 0.05 versus LacZ.