Mouse anti erk

The liver tissue slices were incubated in proteinase K (20 μg/ml) for 5 min for proteolysis and were blocked with 10% normal goat serum for 4 h at room temperature. Then, the samples were incubated for 72 h in a 4℃ humidified chamber with the following primary antibodies: mouse anti-PPARγ (1:50, Santa Cruz, CA), mouse anti-p-mTOR (1:50, Santa Cruz, CA), mouse anti-glutathione peroxidase 4 (GPx4) (1:100, Santa Cruz, CA), mouse anti-8-OHdG (1:100, Santa Cruz, CA), mouse anti-caspase 3 (1:100, Santa Cruz, CA), mouse anti-p-IkB (1:250, Santa Cruz, CA), mouse anti-ERK (1:100, Santa Cruz, CA), and mouse anti-p-JNK0 (1:100, Santa Cruz, CA). Biotinylated goat anti-mouse IgG (1:100, Abcam, Cambridge, UK) secondary antibodies were incubated for 48 h in a 4℃ humidified chamber. Finally, the samples were incubated with the avidin biotin complex solution (ABC kit, Vector Laboratories Inc., Burlingame, CA) for 1 h at room temperature. The immune-reactivity was visualized in a 50-mM Tris−HCl solution (pH 7.4) containing 0.05% 3,3´-diaminobenzidine and 0.01% H₂O₂, followed by counterstaining with hematoxylin.