Detection of the protein expressions of GRP78, CHOP, JNK1, pJNK1 and caspase-12 using Western blot

The total protein of the liver tissue was extracted using the BCA protein extraction method. The same amount of total protein was used to add the loading buffer for denaturation for 3 min. After SDS-PAGE electrophoresis, the proteins were electrotransferred onto the PVDF membrane. After blocking, the proteins were incubated with the corresponding primary antibody (GRP78, CHOP, JNK1, pJNK1 and caspase-12, diluted 1:1,000) at 4°C overnight, respectively. Then, they were incubated with the secondary antibody (diluted 1:7,000) at room temperature for 2 h. After that, ECL was performed, and images were taken using the gel imaging system. Finally, the gray scale scanning analysis was performed for the results.