Extended Data Fig. 3: UTX regulates NK cell fitness.
From: The X-linked epigenetic regulator UTX controls NK cell-intrinsic sex differences
a) Representative flow cytometry plots of splenic NK cells from female WT, male WT, and female UTXHet, and female UTXNKD mice. b) Representative contour plots and c) percent of WT(CD45.1+) or UTXNKD(CD45.2+) cells of total splenic T (left) and NK (right) cells in 1:1 WT:UTXNKD mBMC mice (n = 3). d) Percentage of Ki67+ cells in blood of 4:1 WT:UTXNKD mBMCs (n = 28), injection ratio used to normalize WT and UTXNKD NK cell numbers. e) Representative histogram (left) and quantification (right) of CFSE expansion, division and proliferation indices calculated using FlowJo’s Proliferation tool of CFSE-labeled splenic NK cells isolated from WT and UTXNKD mice stimulated ex vivo with IL-15 (50 ng/mL) for 4 days (n = 4). f) Schematic showing adoptive transfer of CTV-labeled congenically distinct WT (CD451x2) and UTXNKD (CD45.2+) NK cells transferred into WT (CD45.1+) recipients at a 1:1 ratio with analysis of CTV dilution and WT:UTXNKD ratio on D7 by flow cytometry. g) Representative histograms showing CTV dilution of congenically distinct WT (CD451x2) and UTXNKD (CD45.2+) NK cells transferred into WT (CD45.1+) recipients before transfer (left) and on day 7 post-transfer (right). h) Representative histograms and i) percentage of cleaved caspase 3+ splenic NK cells from female WT and male WT treated ex vivo with IL-15 (5 ng/mL) and DMSO (Female WT: n = 7; Male WT: n = 8) or 2.5 uM Nutlin-3a (Female WT: n = 3; Male WT: n = 4) for 24 hours. j) Representative histograms and k) percentage of cleaved caspase 3+ splenic NK cells from gonadectomized female and male mice (n = 6) treated ex vivo with IL-15 (5 ng/mL) and DMSO or 2.5uM Nutlin-3a for 24 h. l) Representative histograms of Bcl-2 (left) and Bim (right) of flow cytometry in splenic NK cells from female WT and UTXNKD mice (n = 5). “Ctrl”:unstained flow cytometry controls. Data are representative of 2-3 independent experiments. Samples were compared using unpaired two-tailed Student’s t test with Welch’s correction and data points are presented as individual mice with the mean ± SEM (N.S., Not Significant; *, p < 0.05; **, p < 0.01; ****, p < 0.0001). Specific p-values are as follows: c:[T cells=0.705; NK Cells=0.0202]; d < 0.0001; e:[Expansion=0.0192; Division=0.0253; Proliferation=0.0032]; i:[DMSO = 0.0096; Nutlin-3a = 0.0473]; k:[DMSO = 0.001; Nutlin-3a = 0.0013].
