Western blotting was performed as previously described [21 (link)]. Cells’ total protein was extracted with lysis buffer (Invitrogen), and the target proteins were separated from total protein by SDS/PAGE. The target proteins on gel were transferred to the PVDF membrane (Millipore). BSA (Gibco) sealed up PVDF membrane for 2 h, proteins were marked by primary antibodies anti-p65 antibody (1:1000, Abcam) or anti-p50 antibody (1:10000, Abcam) or anti-β-actin antibody (1:2000, Abcam) and anti-rabbit HRP secondary antibodies (1:1000, Abcam). PVDF membrane was washed with PBS and we observed the chemiluminescence with ECL Plus Western Blotting Substrate (Thermo Fisher). The film that was placed on the PVDF membrane was exposed and developed.
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