Research Paper Volume 12, Issue 3 pp 2275—2301

PAS kinase deficiency reduces aging effects in mice

Figure 9. Effects of aging and PASK deficiency on the expression of FoxO3a gene and p53, p21, PCNA proteins and the length of telomeres. Real-time PCR was used to analyze the expression of FoxO3a (A) and p53 (B) mRNA levels. The results were measured in the livers of non-fasted (NON-FASTED) and 24-h fasted (FASTED) young (3-5 months) and aged (18-20 months) wild-type (WT) and PASK-deficient (Pask-/-) mice. The mRNA levels of different genes were normalized by the mRNA of β-actin used as housekeeping gene. Immunoblot analysis of p53 (C), p21 (D), PCNA (E) in livers from young and aged WT and Pask-/- mice. Liver lysates from non-fasted (NF) and 24-h fasted (F) mice were processed for western blot analysis. (C, D) The value obtained in 18-20-months-old non-fasted aged WT mice was taken as 1. (E, F) The value obtained in 3-5-month-old non-fasted 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 3) n = 4-5 animals per condition. In addition, telomere length (F) was measured in livers from young and aged WT and Pask-/- mice. $P < 0.05, $$P < 0.01, $$$P < 0.001 3-5 months vs. 18-20 months; * P < 0.05, ** P < 0.01 *** P < 0.001 WT vs. Pask-/-; ##P < 0.01, ###P < 0.001 non-fasted vs. fasted.