Analysis of hepatic expression of β-oxidation modulating genes, Akt and NADH shuttle systems in 70–90-week-old wild-type and AAV mice and hepatic concentrations of NAD+ and NADH. For quantitative RT–PCR, the data represent the mean ± SEM for 70–90-week-old wild-type (n = 22) and AAV (n = 22) mice. (A) Quantification of mRNA for hepatic PPARα, CPT1α, FGF21 and Akt by real-time RT–PCR and quantification of protein levels by densitometric analysis. The data for densitometric analysis represent the mean ± SEM of four separate replicas of western blots. (B) Western-blot analysis of Akt, p-Akt-S473, p-Akt-T308 or β-actin. (C) Quantification of mRNA for members of hepatic MA and GP shuttle systems by real-time RT–PCR. (D) Western-blot analysis of mMDH, mAAT, cMDH, cAAT, mGPDH, cGPDH or β-actin and quantification of protein levels by densitometry of seven separate replicas of western blots. (E) Hepatic levels of NAD+ and NADH in 12-week-old wild-type (n = 12) and AAV (n = 12) mice and 70–90 week-old wild-type (n = 10) and AAV (n = 13) mice. *P < 0.05 and **P < 0.005.