CHD7 interacts with RUNX1, RUNX1/CBFβ, and RUNX1/CBFβ-SMMHC complex. (A-B) 293T cells were transfected with indicated plasmids, and co-immunoprecipitation assays were performed to detect the interaction among these proteins. Flag-tagged CHD7 was immunoprecipitated with anti-Flag M2 beads, and western blot was performed with the indicated antibodies. (A) CHD7 pulls down RUNX1 but not CBFβ or CBFβ-SMMHC. (B) CHD7 pulls down CBFβ and CBFβ-SMMHC in the presence of RUNX1. (C-E) 293T cells were transfected with the indicated plasmids, and immunofluorescence assay was performed to detect the interaction between the indicated proteins. The labels on the left of the photomicrographs indicate the transfected plasmids; labels on the top indicate the observed proteins at the appropriate microscope filter settings. (F) Co-immunoprecipitation assay was performed in ME-1 cells with anti-CHD7 antibody, and western blot was performed with the indicated antibodies. (G) Immunofluorescent staining of leukemic cells from the spleen of an end-stage Mx1-CreCbfb^+/56M mouse to detect the co-localization between RUNX1 and CHD7 (top panel) and between RUNX1 and CBFβ/CBFβ-SMMHC (bottom panel). Scale bars, 10 μm. DAPI, 4′,6-diamidino-2-phenylindole; IgG, immunoglobulin G; IP, immunoprecipitate.