FIGURE 5.

SUMO-1 enhances the association between hDREF and Mi2α. A, 293FT cells were cotransfected with HA-Mi2α (WT or K1971R mutant) with or without CFP-SUMO-1 and FLAG-hDREF plasmids as indicated. At 24 h after DNA transfection, whole cell lysates were prepared and subjected to immunoprecipitation (IP) using anti-HA antibody. hDREF and its SUMOylated form co-immunoprecipitated with Mi2α were detected by immunoblotting analysis (IB) using anti-FLAG antibody. The data are representative of two independent experiments with similar results. B, 293FT cells were cotransfected with HA-Mi2α (WT or K1971R mutant) with and without CFP-SUMO-1 and FLAG-hDREF plasmids as indicated. At 24 h after DNA transfection, whole cell lysates were prepared and subjected to immunoprecipitation using anti-FLAG antibody. Mi2α and its SUMOylated form in the immunoprecipitated material were detected by immunoblotting analysis using anti-HA antibody. The data are representative of two independent experiments with similar results. C, 293FT cells were transfected with FLAG-HDAC1 and HA-Mi2α (WT or K1971R mutant) with or without CFP-SUMO-1 and HA-hDREF plasmids as indicated. At 24 h after DNA transfection, cell lysates were subjected to immunoprecipitation using anti-FLAG antibody. Components of NuRD complex (HDAC1, Mi2α, RbAp48, MTA2, and MBD3) in the immunoprecipitated samples were detected with anti-FLAG, anti-HA antibodies, and specific antibodies against RbAp48, MTA2, and MBD3, respectively. The data are representative of three independent experiments with similar results.