Figure 2. Effects of aging and PASK deficiency on Akt and PTEN protein levels. Immunoblot analysis of total Akt (Akt) (A), phospho-Akt (Ser473) (P-Akt) (B) and PTEN (PTEN) (C) in livers from young (3-5 months) and aged (18-20 months) wild-type (WT) and PASK-deficient (Pask-/-) mice. Liver lysates from non-fasted (NF) and 24-h fasted (F) mice were processed for western blot analysis. The value obtained for non-fasted 3-5-month-old WT mice was taken as 1. Bar graphs represent the means ± SEM of the densitometric values normalized by total protein detected by Stain-Free (TOTAL PROTEIN) (Supplementary Figure 1); n = 4-5 animals per condition. $P < 0.05, $$P < 0.01, $$$P < 0.001 3-5 months vs. 18-20 months; * P < 0.05, ** P < 0.01 WT vs. Pask-/-; #P < 0.05 non-fasted vs. fasted.