Figure S1.

NKG2D is expressed on antigen-experienced CD4⁺ T cells in steady-state C57BL/6 mice. (A) FC gating strategy for the characterization of NKG2D⁺ CD4⁺ T cells in C57BL/6 mice (spleen shown). Lineage (Lin) includes CD19, FcεRI, Gr-1, and F4/80. (B) Representative FC analysis of indicated surface marker expression on NKG2D⁺ (red line) and NKG2D⁻ (black line) CD44⁺ CD4⁺ T cells in the spleen. Shaded histogram represents live CD19⁺ cells (n = 3). (C) Representative FC analysis of DNAM-1 and CD94 expression on splenic CD4⁺ CD44⁺ NKG2D⁻ (black line) and CD4⁺ CD44⁺ NKG2D⁺ (red line). Shaded histogram represents live CD19⁺ cells (n = 6). (D) FC analysis of selected surface marker expression on NKG2D⁺ (red line) and NKG2D⁻ (black line) CD44⁺ CD4⁺ T cells in spleen. Shaded histogram represents NK cells (n = 3). (E) Representative FC analysis of NKG2D expression on CD44⁺ CD4⁺ T cells from spleen of WT, Tyrobp^−/−, Hcst^−/−, and Klrk1^−/− mice. (F) Frequency of NKG2D⁺ CD44⁺ CD4⁺ T cells, quantification of E. Each symbol represents a mouse; line represents the mean ± SEM (n = 3). Data are pooled from two independent experiments. (G) Representative FC analysis of indicated marker expression on NKG2D⁺ and NKG2D⁻ CD44⁺ CD4⁺ T cells in the siLP of Tbx21-ZsGreen mice (n = 3). (H) Representative FC analysis of FoxP3 and CD25 expression on NKG2D⁺ (red) and NKG2D⁻ (gray) CD4⁺ T cells from the spleen of WT mice (n = 3). In A–D, G, and H, data are representative of at least two independent experiments.