Research Paper Volume 12, Issue 3 pp 2275—2301

Figure 6. Effects of aging and PASK deficiency on NRF2, GCLm and HO1 protein expression. Immunoblot analysis of NRF2 (A), GCLm (D) and HO1 (E) 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. Real-time PCR was used to analyze the expression of Gclm (B), Ho1 (C) and Gclc (F) mRNAs. The value obtained in 3-5-month-old non-fasted WT mice was taken as 1. Bar graphs in (A, D, E) represent the means ± SEM of the densitometric values normalized by total protein detected by Stain-Free (TOTAL PROTEIN) (Supplementary Figure 2); (B, C, F) represent the means ± SEM, and the levels of expression were normalized by the mRNA of β-actin, n = 4-5 animals per condition. ^$ P < 0.05, ^$$ P < 0.01 3-5 months vs. 18-20 months; * P < 0.05, ** P < 0.01, *** P < 0.001 WT vs. Pask^-/-; ^# P < 0.05, ^## P < 0.01 non-fasted vs. fasted.