*4.7. Western Blot*

Individual liver of each group for analysis of P65, p-P65, PI3K, p-PI3K, Akt2, p-Akt2, and Glut2 was pulverized, and the proteins were extracted by a total protein extraction kit (Keygen, Shenzhen, China). An equal amount of protein (40 μg) in liver was separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% fat-free milk and incubated with different primary antibodies. The bound antibodies were detected using horseradish peroxidase–conjugated anti-rabbit antibodies. Antibody reactivity was detected by ECL Western blotting Detection Systems.
