Figure 5. mRNA expression levels, Mitochondrial DNA content, Complex I, II and III activities, COX-IV protein content, Cytochrome C Oxidase and Citrate Synthase Activities and mRNA expression levels of Myosin Heavy Chain types in Gastrocnemius of F1 mice.
A) mRNA expression levels of genes encoding proteins involved in the mitochondrial function in Gastrocnemius from 7-months-old F1-CD and F1-LPD males mice. mRNA relative levels are normalized relatively to HPRT1 expression level and expressed as fold change relative to the control value measured in F1-CD. Values are means ± sem for at least 8 mice/group. *p≤0.05. B) Mitochondrial density is estimated by measuring the mitochondria DNA (mtDNA) content relatively to the nuclear DNA (nuclDNA) content in Gastrocnemius from 7-months-old F1-CD and F1-LPD males mice. mtDNA/nuclDNA ratio was calculated as the ratio of COX1 to cyclophilin A DNA levels, determined by real-time PCR, in DNA extracted from gastrocnemius of 7-months-old F1-CD and F1-LPD males mice. Results were normalized by the mean value of the control condition set to 1 unit. *p≤0.05. C) Complex I, II and III and Citrate Synthase (CS) activities are measured in gastrocnemius from 7-months-old F1-CD and F1-LPD males mice and expressed as a percentage of the Cplx/CS ratio of F1-CD mice. Values are means ± sem for at least 8 mice/group. D) Left panel: Cytochrome C Oxidase Sub-Unit IV (COX-IV) protein level was assessed by western blotting in gastrocnemius from 7-months-old F1-CD and F1-LPD males mice. Histograms shown represent a densitometrical quantification after Porin normalization. Values are means ± sem for at least 4 mice/group. *p≤0.05. Right panel: Cytochrome C Oxidase (COX) and Citrate Synthase (CS) activities are measured in gastrocnemius from 7-months-old F1-CD and F1-LPD males mice and expressed as a percentage of the COX/CS ratio of F1-CD mice. Values are means ± sem for at least 8 mice/group. *p≤0.05. E) mRNA expression levels of the 4 different types of Myosin Heavy Chain (MHC-I, MHC-IIa, MHC-IIb, MHC-IIx) in Gastrocnemius from 7-months-old F1-CD and F1-LPD males mice. mRNA relative levels are normalized relatively to HPRT1 expression level and expressed as fold change relative to the control value measured in F1-CD. Values are means ± sem for at least 8 mice/group. *p≤0.05.