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. 2021 Apr 15;218(6):e20202032. doi: 10.1084/jem.20202032

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© 2021 Li et al.

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Figure 4. — TCF1 deficiency in mature NK cells impacts NK cell receptor expression and NK cell function. (A) Flow cytometry plots of multiple surface proteins on BM NK cells from Ctrl (white) and Tcf7^Δ/Δ (maroon) mice. The percentage of cells in the gated region is indicated (n = 3 or 4 for each group in two to three independent experiments). (B) Summary of data shown in A. MFI of Ctrl NK cells was set as 1 in each experiment. (C) CD49b⁺-enriched NK cells from BM were stimulated by IL-2 and IL-12 followed by analysis of IFN-γ in NK1.1⁺CD49b⁺ NK cells by flow cytometry. Numbers are the percentage of cells in the indicated gates (n = 3). (D) Summary of data shown in C. (E) Flow cytometry for IFN-γ in NK1.1⁺CD49b⁺ NK cells 18 h after activation with anti-NK1.1 (n = 3). (F) Summary of data shown in E. (G) C57BL/6 (H-2^b) and β2m^−/− splenocytes were labeled with a low and high dose of CFSE, respectively, before injection into polyinosinic:polycytidylic acid–primed Tcf7^Δ/Δ and Ctrl mice. Recipient spleens were analyzed 18 h later by flow cytometry. Numbers are the percentage of cells in the indicated gates (n = 3). (H) Summary showing percent rejection in Ctrl and Tcf7^Δ/Δ mice (G). (I) Ctrl and Tcf7^Δ/Δ mice were injected with B16-F10 cells, and lungs were analyzed for metastases on day 12 (n = 3 for each group in two independent experiments). (J) Summary of data shown in I for numbers of tumor nodules in the lung. Note that there is no significant difference between Ctrl and Tcf7^Δ/Δ mice; Id2^Δ/Δ mice are also shown for comparison. Error bars represent SEM (B, D, F, H, and J). Statistical significance was determined by two-tailed unpaired t test (B, D, F, H, and J). *, P < 0.05; **, P < 0.01. SSC, side scatter.