Coimmunoprecipitation (Co-IP)
The cells were washed three times with PBS and subsequently lysed on ice
in lysis buffer (Beyotime, Beijing, China). Next, the lysates were
centrifuged at 12,000 × g for 15 min. Subsequently, the cell lysates
were incubated with specific antibodies for 1 h, followed by a 30-minute
incubation with Dynabeads™ Protein G beads at 4°C. The
beads were then washed three times with cold lysis buffer. Following the
washes, the beads were boiled in 10 µL of 5× loading buffer (Beyotime,
Shanghai, China) for 5 min before being analyzed by western blot using
antibodies against DNMT1, G9a, E2F1, Flag, Myc, and GST.