Leukemia cells were resuspended in glucose-free DMEM and incubated with NBDG at 37°C in the dark for 30 minutes at 1ul of 5mg/ml NBDG per 3 million cells (27 (link)). After 30 minutes of incubation, leukemic cells were washed with PBS. The cells were then stained with anti-HLA antibody and 4’, 6-Diamidino-2-Phenylindole, Dilactate (DAPI) (Thermo Fisher Scientific). NBDG-low, NBDG-med, or NBDG-high cells were sorted by Cytopeia InFlux Cell Sorter (BD Biosciences) and BD Influx Cell Sorter (BD Biosciences) using 488nm excitation and 505-522nm emission wavelengths.
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