Mi2β is modified by O-GlcNAc.
a, O-GlcNAcylated protein was immunoprecipitated by an O-GlcNAc-specific antibody (CTD 110.6) in K562 cells before and after NaB induction (top panel) or H-H induction (bottom panel); the blot was probed with Mi2β antibody. b, Mi2β protein was immunoprecipitated from K562 cells before and after NaB induction (top panel) or H-H induction (bottom panel); the blot was probed with CTD 110.6. c and d, Mi2β protein was immunoprecipitated from K562 cells, and the blot was probed with CTD 110.6 (c) or CTD 110.6 preincubated with 500 mm GlcNAc (d). e, Mi2β protein was immunoprecipitated from K562 cells and treated with hexosaminidase CpNagJ at 37 °C for 3 h; the blot was probed with CTD 110.6. The OGA inhibitor TMG was used to increase overall O-GlcNAc levels. Isotype immunoprecipitation (normal mouse IgM or rabbit IgG) and antibody-alone immunoprecipitation (1°) were used as negative controls. All experiments were performed with at least three biological replicates. WB, Western blotting.