To determine whether the NF-κB pathway is elevated in human NPmut AML cells, we quantified the nuclear versus cytosolic localization of NF-κB p65 in K562 and KY821 cells with or without 3 ng/mL TNF-α stimulation using a confocal immunofluorescence microscopy–based method similar to that described in a previous publication (50) (Figure 6E). Here, NFKB1 is linked to acute myeloid leukemia.