FIGURE 5.

HDAC1 and -2 maintains the interactions with corepressor complex proteins during mitosis. A and B, total cell lysates (500 μg) from cycling and nocodazole (Noc)-treated HeLa cells were incubated with 3.0 μg of rabbit anti-HDAC1 (A) or anti-HDAC2 (B) antibodies. The immunoprecipitated fractions were checked with antibodies against Sin3A, RbAp48, or CoREST. Isotype-specific non-related rabbit IgG was used as negative control. IP and ID, immunoprecipitated and immunodepleted fractions, respectively. C, total cellular lysates (500 μg) from cycling (untreated) and nocodazole-treated (mitotic) HeLa cells were incubated with 3.0 μg of mouse anti-RbAp48 antibody. The immunoprecipitated fractions were analyzed by anti-HDAC1 and anti-HDAC2 antibodies. Isotype-specific non-related mouse IgG was used as negative control. IN, IP, and ID represent input, immunoprecipitated, and immunodepleted fractions, respectively. The slower migrating band in the RbAp48 immunoprecipitated fraction may be phosphorylated RbAp48, but this has not been validated. The representative immunoblots are shown from one of three independent experiments, which are used for quantifications as mentioned under “Experimental Procedures.”