Nuclear or cytoplasmic proteins (20 μg) were loaded onto each well, separated on 10% SDS-polyacrylamide gel and electroblotted onto nitro-cellulose membranes. Nonspecific binding was blocked by incubation of the membranes in TBS/1% nonfat dried milk/0.1% Tween 20 followed by Abs indicated in the figure legends. The Abs against phospho and total Erk, JNK, and p38 were purchased from Cell Signaling Technology and CYP2E1 and IRAK-M was from Chemicon. The rabbit Ab against IκBα was from Santa Cruz Biotechnology. β-actin and TATA-binding protein Abs were from Abcam. The Abs were detected using HRP-conjugated secondary Abs (Santa Cruz Biotechnology) and chemiluminescence assay reagents from Cell Signaling Technology.
