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. 2014 Aug 13;9(8):e104896. doi: 10.1371/journal.pone.0104896

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.

Figure 5

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.