Zhx2 inhibits NK cell functions. (A) Percentage of indicated effector molecules assessed with FCM in anti-NK1.1–stimulated CD27⁻CD11b⁺ liver NK cells. Cells were pregated on CD3⁻NK1.1⁺ subsets (representative of three independent experiments). (B) Percentage of indicated effector molecules assessed with FCM in anti-NK1.1–stimulated CD27⁻CD11b⁺ spleen NK cells. Cells were pregated on CD3⁻NK1.1⁺ subsets (representative of three independent experiments). (C) IFN-γ, TNF-α, perforin, and granzyme production in NK92 cells transfected with ZHX2 overexpression (ZHX2-OE) plasmid or empty vector (control; representative of three independent experiments). (D) IFN-γ, TNF-α, perforin, and granzyme production in NK92 cells infected with ZHX2 shRNA expressing lentivirus (shZHX2) or control shRNA lentivirus (representative of three independent experiments). (E) NK92 cells with indicated treatment were cocultured with CFSE-labeled K562 cells for 6 h. FCM assay was used to detect 7-AAD staining in K562 cells (representative of three independent experiments). Each symbol represents data from an individual sample, 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 < 0.0001; ***, P < 0.001; ****, P < 0.0001.