Western blot analysis
Cells were harvested after treatment for 24 h, and total proteins were
extracted with RIPA lysis buffer
(Beyotime).
After separation using 12% SDS-polyacrylamide gels, the proteins were
transferred onto PVDF membranes (Millipore, USA). The PVDF membranes
were blocked with Quick Blocking Buffer (Beyotime) for 15 min and then
incubated overnight with primary antibodies at 4 °C. The antibodies
described above were used. After incubation for 1 h with secondary
antibodies in an incubator at 37 °C, the signals were identified by
chemiluminescence detection.