EPZ004777 treatment induces apoptosis, cell-cycle arrest, and terminal differentiation in DNMT3A-mutant human AML cells. HL60, MV411, OCI AML2, and OCI AML3 cells were treated with 3 μM EPZ004777 or DMSO vehicle control for 14 days. Cells were replated at a constant concentration in fresh drug-containing media every 2 to 3 days. (A) Representative flow cytometry histograms of AVB for OCI AML2 and OCI AML3 cells on day 14 of treatment with DMSO vehicle control (blue) or EPZ004777 (red) (left). Quantification of % of EPZ004777-treated cells that are AVB+ minus % AVB+ vehicle control–treated cells (right). (B) Representative flow cytometry plots of propidium iodide (PI) DNA content cell-cycle analysis for OCI AML2 cells and OCI AML3 cells (left) with quantification of experiments (right) (C) Representative flow plots of CD14 cell surface expression of OCI AML 2 and OCI AML 3 cells treated with DMSO vehicle control or EPZ004777 for 14 days after gating out PI+ dead cells (left) and quantification of percentage of CD14+ cells treated with 3 μM EPZ004777 or vehicle control at specified time points (right). All assays were done in triplicate. Error bars represent standard deviation. SSC, side scatter.