Cell cycle assay

AML cell lines were seeded in a 48-well plate at a density of 0.25 × 10⁶ cells/well and incubated with increasing concentrations of CPI-637, NEO1132, or NEO2734 for 24 and 48 hours. Cells were fixed in ice-cold 70% ethanol, incubated for 1 hour at 4°C, and washed twice with phosphate buffered saline (PBS). Subsequently, cells were stained with 50 µg/mL propidium iodide (Sigma Aldrich) in PBS supplemented with 100 µg/mL RNAse A (Sigma Aldrich) and analysis was performed using a FACS-Fortessa flow cytometer (BD Biosciences). Data analysis was performed with FACS Diva Software (BD Biosciences).