4.7. Western Blot Analyses

Total proteins of whole cell lysates or cytoplasmic and nuclear extracts were assayed using Bradford’s reagent (Bio-Rad, Hercules, CA, USA). The proteins were divided using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose (NC) membrane (Millipore, Bedford, MA, USA). After blocking of nonspecific sites with 3% BSA, membranes were incubated with each primary antibody at room temperature (RT) for 2 h or at 4 °C overnight. The membranes were subsequently incubated with HRP-conjugated anti-mouse or anti-rabbit secondary antibodies. Signals were detected using Super Signal West Femto chemiluminescent substrate (Thermo Scientific, Rockford, IL, USA). Protein levels were quantified using a ChemiDoc^TM Touch Imaging System (Bio-Rad, Hercules, CA, USA).