Extended Data Fig. 4: UTX enhances effector function independent of gonadal hormone and maturation.
From: The X-linked epigenetic regulator UTX controls NK cell-intrinsic sex differences
a) Representative contour plots of total NK cells from female WT, male WT, female UTXHet, and female UTXNKD mice cultured with IL-15 (50 ng/mL) and IL-12 (20 ng/mL) for 4 h. b) Absolute number of IFN-γ+ NK cells from female WT, male WT, and female UTXHet mice stimulated with no treatment (NT) or IL-15 (50 ng/mL) and IL-12 (20 ng/mL) for 4 h (n = 8). c) Specific lysis of MHC Class I deficient MC38 (Target) cells by female WT (n = 4), male WT (n = 5), and gonadectomized female (n = 3) and male (n = 6) NK cells for 16 h at 4:1 effector:target ratio, normalized to lysis by female WT. d) Representative histograms and e) MFI of CD107a (n = 5 per group), granzyme b (GzmB) (Female:n = 5; Male:n = 4), and perforin (n = 5) of female WT and male WT NK cells incubated with IL-15 (50 ng/mL) only or additionally stimulated with plate-bound anti-NK1.1 antibody (PK136). “Ctrl” refers to matched unstained control for flow cytometry. Representative f) contour plots of IFN-γ and g) histogram of GzmB expressing total splenic NK cells on D1.5 post MCMV infection of 4:1 WT:UTXNKD mixed bone marrow chimeras (mBMCs), ratio used to normalize cell numbers between genotypes. h) IFN-γ protein production in UTXNKD compared to WT NK cells within maturation subsets: CD27−CD11b− (DN), CD27+CD11b− (CD27 SP), CD27+CD11b+ (DP), and CD27−CD11b+ (CD11b SP) isolated from 4:1 WT:UTXNKD mBMC 1.5 days post-MCMV infection (n = 6). i) Representative contour plots of total NK cells derived from 1:1 WT:iUTX-/- mBMC mice on day 1.5 post MCMV infection, normalized to WT (n = 6). Data are representative of 2-3 independent experiments. Samples were compared using paired two-tailed Student’s t test and data points are presented as individual mice with the mean ± SEM (N.S., Not Significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001). Specific p-values are as follows: b:[F WT vs. M WT = 0.0162; M WT vs. UTXHet = 0.968; F WT vs. F UTXHet = 0.0288]; c:[F WT vs. F Gonadectomy=0.245; F WT vs. M WT = 0.0086; M WT vs. M Gonadectomy=0.998; F Gonadectomy vs. M Gonadectomy=0.0247]; e:[CD107a – NT = 0.54 and NK1.1 = 0.772; GzmB – NT = 0.0004 and NK1.1 = 0.00129; Perforin – NT = 0.0101 and NK1.1 < .0001]; h:[DN = 0.03281; CD27 SP = 0.0186; DP = 0.0231; CD11b SP = 0.0114].
