Figure 2.

Acute deletion of Atg7 enhances alcohol-induced liver injury in a chronic-plus-binge model. A: Scheme of the chronic-plus-binge treatment. Mice were acclimated with liquid diet for 5 days. Atg7^F/F and Atg7^Δhep-ERT2 mice were injected with tamoxifen (TMX) on days 3 and 4 during acclimation. After acclimation, mice were randomly divided into two groups and given a liquid diet with alcohol (EtOH-fed) or with maltose dextrin (Pair-fed) for 10 days, followed by a single gavage of alcohol (5 g/kg) or isocaloric maltose dextrin. Mice were analyzed 9 hours later. B: Representative Western blot images of protein expression in the livers of Atg7^F/F and Atg7^Δhep-ERT2 mice after alcohol or pair-fed treatment. Asterisk indicates onspecific bands. C: Densitometry of each protein band was conducted. The density values of Atg7, p62, or Nad(p)h quinone dehydrogenase (Nqo)-1 were normalized to that of β-actin. The density of microtubule-associated protein 1A/1B-light chain (Lc)-3II was normalized to that of Lc3I. D: The mRNA levels of Gstm1 and Nqo1 in the livers of Atg7^F/F and Atg7^Δhep-ERT2 mice after alcohol or pair-fed treatment. Expression levels were normalized to that of β-actin. E and F: Liver weight and body (L/B) weight ratio, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bile acids (TBA) in Atg7^F/F and Atg7^Δhep-ERT2 mice after alcohol or pair-fed treatment. G: Hepatic triglycerides (TG) and serum β-hydroxybutyrate (β-OHB) levels in Atg7^F/F and Atg7^Δhep-ERT2 mice given alcohol or pair-fed treatment. Mice were used at the age of 8 to 12 weeks. Data are expressed as the means ± SEM fold changes over those of pair-fed Atg7^F/F mice. n = 4 (A, C, and D); n = 4 to 11 (G). ^∗P < 0.05 versus nontreated F/F control; ^†P < 0.05 versus nontreated Δhep control; ^‡P < 0.05 versus treated F/F control. Gstm1, glutathione S-transferase M1; and Nqo, Nad(p)h quinone dehydrogenase 1; Sqstm, sequestosome.