2.4 |. Apoptosis assay

After 72-hours of drug treatment, CLBL-1 cells treated with DMSO or pevonedistat at concentrations of 0.25, 0.5 and 1.0 μM were collected and were stained with FITC-conjugated Annexin V (1:40; eBioscience, Inc., San Diego, CA) and SYTOX DNA binding dye (1:1000; Thermo Fisher Scientific, Madison, WI). Primary DLBCL samples treated with DMSO or pevonedistat (2.0 μM) were harvested after 12 hours of treatment and stained with fluorescein isothiocyanate (FITC)-conjugated Annexin V (1:40; eBioscience, Inc., San Diego, CA) and DAPI (1:500; Thermo Fisher Scientific, Madison, WI) and were subjected to evaluation by flow cytometry. Samples were analysed using BD LSRII flow cytometry machine (BD Biosciences, San Jose, CA), and results were analysed using FlowJo v10.0.7 software.