Western blot analysis
The treated cells were harvested and lysed in RIPA buffer. In some experiments, NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher, USA) were used to isolate the nuclear and cytosolic proteins according to the manufacturer’s instructions. Equal amounts of proteins from each group were loaded onto a 10% SDS-PAGE gel. After electrophoresis, the proteins were transferred to membranes, and the membranes were blocked with 5% non-flat milk and incubated with primary antibodies overnight at 4ºC. Subsequently, the proteins were incubated with the corresponding HPR-conjugated secondary antibodies for 30 min at 37ºC and visualized and imaged using a chemiluminescence (ECL) detection system.