Knockout of Zhx2 in NK cells results in increased maturation of NK cells. (A and B) FACS plots and bar graphs present percentages and absolute numbers of CD3⁻NK1.1⁺CD49b⁺ NK cells in the liver, spleen, and blood from Zhx2^+/+Ncr1-icre⁺ (Zhx2^+/+) and Zhx2^fl/flNcr1-icre⁺ (Zhx2^Δ/Δ) mice (representative of at least three independent experiments). (C–E) FACS plots (C) and bar graphs of cell percentage (D) and cell number (E) depict NK cell subsets determined by expression of CD11b/CD27 in the liver, spleen, and blood from Zhx2^+/+ and Zhx2^Δ/Δ mice (pregated on CD3⁻NK1.1⁺CD49b⁺ NK subsets, representative of at least three independent experiments). DN, double negative. (F) Representative histograms showing KLRG1 expression in liver and spleen CD3⁻NK1.1⁺CD49b⁺ NK cells from Zhx2^+/+ and Zhx2^Δ/Δ mice. Graphs show the quantification of the percentage of KLRG1⁺ NK cells (representative of three independent experiments). FSC, forward scatter. (G) Graphical representation of in vitro NK cell differentiation system using NK cells isolated from Cag-Zhx2^+/+ and Cag-Zhx2^Δ/Δ mice. (H) Representative plots of expression of CD11b/CD27 in gated CD3⁻NK1.1⁺ cells and graph depicting percentages of CD27⁻CD11b⁺ cells (representative of two independent experiments). For all figures, dots represent individual mice or different cell replicates, and error bars represent SEM per group in one experiment. Data were analyzed using Student’s t test (two-tailed paired t test). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.