2.11 | Western blot
Protein lysates from tissue and cell sources were separated by SDS-PAGE, and transferred to PVDF membranes at 4 °C. The blots were washed with TBST (TBS + 0.1% Tween-20), blocked with 7% skim milk in TBST for 1 h, and incubated with the specific primary antibodies overnight at 4 °C on a shaking table. Next, the membranes were incubated with HRP-conjugated anti-mouse or anti-rabbit IgG secondary antibodies (1:5000) in 2% skim milk for 2 h with shaking at room temperature. The immunoblots were visualised with an ECL kit (Zen-Bio, Inc., China).