Protein Extraction and Western Blotting

Cytoplasmic and nuclear proteins were extracted from cells using BeyoECL Plus Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime) supplemented with protease inhibitors and phosphatase inhibitors. Western blotting was performed as previously described [11 (link)]. Primary antibodies specific to GAPDH (1:1000 dilution, Cell Signaling Technology, Danvers, Massachusetts, USA), H3 (1:1000 dilution, Cell Signaling Technology), and APC (1:1000 dilution, Abcam, Cambridge, UK) were used. The blots were then incubated with goat anti-rabbit or anti-mouse secondary antibody (1:3000 dilution, Abcam) and visualized by Immobilon Western Chemiluminescent HRP Substrate (Millipore, St. Louis, MO, USA).