Western Blot Analysis of Protein Samples

The protein samples were prepared from the cell pellets as previously described [18 (link),19 (link)]. Then the samples were separated on SDS-PAGE gel and transferred to NC (nitrocellulose, NC) membrane. After blocking in 5% nonfat milk, primary antibodies were added and incubated with membrane overnight at 4 °C (anti-Flag (1:5000, M2, Sigma), anti-DNMT3B (1:1000), anti-actin (1:10,000, clone C4, Sigma)). Secondary antibody was incubated for 1 h at room temperature (goat anti-mouse HRP (1:5000, Abcam), goat anti-rabbit (1:10,000, Abcam)). The images of Western blot were captured by image lab version 5.1 (Bio-Rad Laboratories, Hercules, California, USA).