Fig. 1. HK2 localizes to the nuclei of leukaemic stem and progenitor cells.
a, Immunoblot of glycolytic and tricarboxylic acid-cycle enzymes in the nucleus, cytoplasm and whole-cell lysate of FACS-sorted stem and bulk 8227 cells. Representative immunoblot from n = 3 biological repeats. b, Confocal microscopy images of HK2 and the mitochondrial protein Tom20 in FACS-sorted stem and bulk 8227 cells. Representative images from n = 3 biological repeats. c, Confocal microscopy images of HK2 in ROS-low LSCs and ROS-high bulk primary cells from patients with AML. Images are representative of three biologically independent samples. d, Nuclear HK2 expression in samples from patients with AML (n = 25) and AML cell lines (n = 15), determined using RPPA. Patient samples: minimum, −2.696; maximum, −1.200; and median −1.679; AML cell lines: minimum, −3.1878; maximum, −0.5461; and median, −1.997. In the box-and-whisker plots, the horizontal lines mark the median, the box limits indicate the 25th and 75th percentiles, and the whiskers extend to 1.5× the interquartile range from the 25th and 75th percentiles. e, 8227 cells were transduced with NLS1–HK2 or control, using a blue fluorescent protein (BFP)-expressing vector. BFP-sorted cells were imaged using confocal microscopy. Representative images of HK2 in control-vector and NLS1–HK2 8227 cells from n = 3 biological repeats are shown. f, The right femur of NOD/SCID-GF mice (n = 7 EV and 8 NLS1–HK2 mice) was injected with 8227 cells transduced with NLS–HK2 or control vector. Eight weeks post injection, engraftment of 8227 cells into the uninjected left femur was measured by flow cytometry. g, Cells from f were injected into secondary mice and the engraftment efficiency was measured 8 weeks later by flow cytometry (n = 7 mice per group). b,c,e, Scale bars, 10 µm. f,g, Statistical analyses were performed using a two-tailed unpaired Student’s t-test. Data represent the mean ± s.e.m.