1, Table 1). Intracellular macroscopic lipid according to fat score increased in both ethanol-fed groups (Table 1). There were nonsignificant increases in inflammatory cells and necrosis in the heterozygote ethanol-fed group and no fibrosis in any mice. TUNEL assay revealed increased hepatocellular apoptosis in both genotype and ethanol feeding, with additive effects in the Het-E group (Table 1). Liver GSH, a measure of antioxidant defense capacity, was reduced in the heterozygous control and in both ethanol-fed

groups, with additive effects of ethanol feeding and genotype in the Het-E group (Table Linsitinib mouse 1). There were no differences among the groups in liver homocysteine levels. Liver SAM was reduced and SAH was elevated in both

ethanol-fed groups, with an additive effect of genotype on SAH in the Het-E group. The SAM/SAH ratio of methylation capacity decreased in both ethanol fed groups, with interactive effects of genotype and ethanol in the Het-E group. The SAM/SAH ratio correlated negatively with the total pathology score (r = −0.57, P < 0.006) and TUNEL score (r = −0.52, P < 0.01). Scatter plots of these DNA Damage inhibitor and subsequent regression analyses are shown in Supporting Figs. 1–5. ER chaperone GRP78 messenger RNA (mRNA) (Table 2) and its protein levels (Fig. 2A) increased in both ethanol-fed groups, with an interaction of genotype and ethanol on protein levels in the Het-E group. Protein levels of the ER stress transducer ATF4 increased in both ethanol groups with greatest and interactive 上海皓元 effects in the Het-E group (Fig. 2B). Activated ER stress transducer ATF6 increased by genotype and maximally in the Het-E group (Fig. 3C). Liver transcript levels of the pro-apoptotic

gene GADD153 increased in both ethanol-fed groups (Table 2), while protein expression rose with both genotype and ethanol feeding, with interactive effects in the Het-E group (Fig. 2D). Cleaved caspase 12, a protease that plays a central role in initiating ER stress-induced apoptosis, increased in both groups of ethanol-fed mice (Fig. 2E). Transcript and protein levels of SREBP-1c increased in both ethanol-fed groups, with additive and interactive effects of both treatments on mRNA expression in the Het-E group (Table 2, Fig. 2F). Ethanol feeding increased SREBP-1c targeted transcripts of acetyl-coenzyme A carboxylase, with interactive effects in the Het-E group, while fatty acid synthase expression rose by genotype only (Table 2). The SAM/SAH ratio of methylation capacity correlated negatively with protein levels of GRP78 (r = −0.43, P < 0.04), GADD153 (r = −0.62, P < 0.002), and cleaved caspase-12 (r = −0.73, P < 0.002). The percentages of methylated cytosine were similar among all groups: 4.01% ± 0.03 in wild-type controls, 4.0% ± 0.1 in heterozygous controls, 3.8% ± 0.01 in wild-type ethanol-fed, and 3.9% ± 0.2 in Het-E mice.