The liver tissue was homogenized as described previously.31 (link), 32 (link) Total liver lysates were then used to quantify GRP78, CHOP, total PERK and pPERK, total and peIF2α, ATF4, ATF6α, ATF6β, sXBP-1, TRAF2, cleaved caspase 3, caspase 12, cytochrome c, total and phospho-ERK, total and phospho-p38 MAPK, total and phospho-GSK3β, total and phospho-JNK, total and phospho-AKT, as well as total and phospho-VDAC by western blot. Cytosolic fractions were used to quantify cleaved caspase 9 and cytochrome c by western blot. Proteins were separated by SDS-PAGE and transferred into polyvinylidene fluoride membranes. Membranes were immunoblotted with antibodies directed against GRP78, CHOP, total and pPERK, total and peIF2α, ATF4, ATF6α, ATF6β, sXBP-1, β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and TRAF2, cleaved caspase 3, cleaved 9 and caspase 12, cytochrome c, total and phospho-ERK, total and phospho-p38 MAPK, total and phospho-GSK3β, total and phospho-JNK and total and phospho-AKT (Cell Signalling Technology Inc., Beverly, MA, USA). The bands were visualized using an enhanced chemiluminescence kit (Bio-Rad Laboratories, Hercules, CA, USA). The values were obtained by densitometric scanning and the Quantity One software program (Bio-Rad Laboratories, Hercules, CA, USA). The scanning values for GRP78, CHOP, ATF4, ATF6α, ATF6β, sXBP-1, TRAF2, cleaved caspase 3, cleaved caspase 9, caspase 12 and cytochrome c were divided by the scanning values for β-actin, and those for phosphorylated PERK, eIF2α, GSK3β, VDAC, ERK, JNK and p38 MAPK by the total PERK, eIF2α, GSK3β, VDAC, ERK, JNK and p38 MAPK, respectively.