Figure 3. Chronic APAP administration at an infratherapeutic dose increases Nrf2-targets HO-1, NQO1 and MnSOD and enhanced antioxidant capacity. (A) Representative Western blots showing HO-1, MnSOD and NQO1 protein levels in liver extracts from 2 m/o, 14 m/o and 14 m/o + APAP mice. GAPDH and VINCULIN levels were used as loading controls (n = 6-14 mice per group) (see original western blot in Supplementary Figure 6). (B) Liver mRNA expression of Nfe2l2, Hmox1, Sod2 and Nqo1 from 2 m/o, 14 m/o and 14 m/o + APAP mice, analyzed by qRT-PCR. Values have been normalized against Hprt1 mRNA, and expressed as fold of change vs. 2 m/o (n = 6-8 mice per group). (C) Hepatic LPO levels of 2 m/o, 14 m/o and 14 m/o + APAP mice (n = 6-8 mice per group). (D) Detection of O2•− in frozen liver sections from 2 m/o, 14 m/o and 14 m/o + APAP group mice using DHE (n = 5 mice per group). (E) Representative Western blots showing phospho-ERK1/2 and phospho-p38 MAPK protein levels. Total ERK1/2 and p38 MAPK levels were used as loading controls, respectively (n = 9-14 mice per group) (see original western blot in Supplementary Figure 7). For (A, E), graphs depict densitometric quantifications of the indicated protein levels. Data are represented as the mean ± S.E.M. Statistical analysis was performed by Kruskal-Wallis, one-way ANOVA or Brown-Forsythe and Welch ANOVA test followed by their respective post-hoc test. * P < 0.05 and *** P < 0.001 vs. 2 m/o; #P < 0.05 and ###P < 0.001 vs. 14 m/o.