(A) Effect of 1 μM AI-10-49 on CBFβ-RUNX1 and CBFβ-SMMHC-RUNX1 binding at 3 and 6 hours in ME-1 cells, measured by coimmunoprecipitation (quantification of three experiments is shown on the right). (B) Schematic of the effect of CBFβ-SMMHC on RUNX1 occupancy and target gene expression and the effect of AI-10-49 on occupancy and expression. (C) Chromatin immunoprecipitation assay showing RUNX1 occupancy on RUNX3, CSF1R, and CEBPA in ME-1 and U937 cells treated with 1 μM AI-10-49 for 6 hours and represented as fold enrichment relative to DMSOtreated cells. Each symbol represents the mean of triplicate experiments; error bars represent the SD. (D) Relative expression (qRT-PCR) of RUNX3, CSF1R, and CEBPA in ME-1 and U937 cells treated with 1 μM AI-10-49 for 6 and 12 hours, and normalized to the DMSO control group. Each symbol represents the mean of triplicate experiments; error bars represent the SD. For all panels, significance was calculated as unpaired t-test, *P < 0.05, or ***P < 0.001.