Zhx2 deficiency gives NK cells enhanced effector functions and promotes tumor immunosurveillance. (A) Representative plots showing the expression of NKG2A and NKG2D in spleen NK cells from Zhx2+/+ and Zhx2Δ/Δ mice. Graphs show the quantification of percentages of NKG2A+/NKG2D+ NK cells (representative of three independent experiments). FSC-H, forward scatter height. (B) Percentages of indicated effector molecules accessed with FCM in anti-NK1.1–stimulated NK cells from the liver and spleen of Zhx2+/+ or Zhx2Δ/Δ mice (representative of three independent experiments). (C) Real-time qPCR analysis of expression of granzyme, IFN-γ, and TNF-α in splenic NK cells from Zhx2+/+ or Zhx2Δ/Δ mice (representative of two independent experiments). (D and E) FACS analysis of the cytotoxicity of the indicated NK cells against CFSE-labeled Yac-1 cells. D shows the relative lysis of Yac-1 cells detected by 7-AAD staining at different E:T ratios (representative of three independent experiments). E shows real-time cytotoxicity assays against CFSE-labeled Yac-1 cells at 5:1 E:T ratio. Yac-1 cell growth was measured with the Real-Time Cell Analyzer multiple plate system (PerkinElmer; representative of three independent experiments). MFI, mean fluorescence intensity. (F) In vivo killing assays were performed by i.p. injection of CFSE-labeled Yac-1 cells in the indicated mice. Representative plots and frequencies and numbers of remaining CFSE-labeled Yac-1 cells in the peritoneal cavity are displayed (representative of two independent experiments). (G and H) Xenograft assays in Zhx2+/+ and Zhx2Δ/Δ mice (representative of two independent experiments). G shows ex vivo tumor imaging. Scale bar: 1 cm. H depicts the tumor weight and growth of s.c. inoculated Hepa1-6 cells in the indicated mice. Each dot represents data from an individual mouse, and error bars represent SEM per group in one experiment. Data were analyzed using Student’s t test (two-tailed paired t test) for A–C, F, and H (left), and using two-way ANOVA for D, E, and H (right). *, P < 0.05; **, P < 0.01; ***, P < 0.001
Figure 3.

Zhx2 deficiency gives NK cells enhanced effector functions and promotes tumor immunosurveillance . (A) Representative plots showing the expression of NKG2A and NKG2D in spleen NK cells from Zhx2+/+ and Zhx2Δ/Δ mice. Graphs show the quantification of percentages of NKG2A+/NKG2D+ NK cells (representative of three independent experiments). FSC-H, forward scatter height. (B) Percentages of indicated effector molecules accessed with FCM in anti-NK1.1–stimulated NK cells from the liver and spleen of Zhx2+/+ or Zhx2Δ/Δ mice (representative of three independent experiments). (C) Real-time qPCR analysis of expression of granzyme, IFN-γ, and TNF-α in splenic NK cells from Zhx2+/+ or Zhx2Δ/Δ mice (representative of two independent experiments). (D and E) FACS analysis of the cytotoxicity of the indicated NK cells against CFSE-labeled Yac-1 cells. D shows the relative lysis of Yac-1 cells detected by 7-AAD staining at different E:T ratios (representative of three independent experiments). E shows real-time cytotoxicity assays against CFSE-labeled Yac-1 cells at 5:1 E:T ratio. Yac-1 cell growth was measured with the Real-Time Cell Analyzer multiple plate system (PerkinElmer; representative of three independent experiments). MFI, mean fluorescence intensity. (F) In vivo killing assays were performed by i.p. injection of CFSE-labeled Yac-1 cells in the indicated mice. Representative plots and frequencies and numbers of remaining CFSE-labeled Yac-1 cells in the peritoneal cavity are displayed (representative of two independent experiments). (G and H) Xenograft assays in Zhx2+/+ and Zhx2Δ/Δ mice (representative of two independent experiments). G shows ex vivo tumor imaging. Scale bar: 1 cm. H depicts the tumor weight and growth of s.c. inoculated Hepa1-6 cells in the indicated mice. Each dot represents data from an individual mouse, and error bars represent SEM per group in one experiment. Data were analyzed using Student’s t test (two-tailed paired t test) for A–C, F, and H (left), and using two-way ANOVA for D, E, and H (right). *, P < 0.05; **, P < 0.01; ***, P < 0.001

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