Rats were allowed free access to an ethanol-containing diet or pair-fed a control diet for 4 and 7 days. (A) Formalin-fixed subcutaneous adipose sections were stained with an antibody against 4-HNE protein adducts. (B) Immunoreactive CYP2E1 was measured in subcutaneous adipose tissue homogenates by Western blotting. ERK1/2 was used as loading control. The values (means ± SEM, arbitrary units of density for CYP2E1/ERK1/2, n=4) were 0.16 ± 0.04 for pair-fed, 0.40 ± 0.08 at day 4 and 0.57 ± 0.14 at day 7. CYP2E1 expression was increased at day 7 compared to pair-fed, p<0.05. (C) XBP-1 splicing was detected by RT-PCR. (D) Immunoreactive phospho-eIF2α, CHOP and grp78 in subcutaneous adipose tissue homogenates were measured by Western blotting. ERK1/2 was used as loading control. Images are representative of 3 (A) and 4–6 (B, C and D) rats in each experimental group. P: pair-fed, E: ethanol-fed, Tg: thapsigargin.