CD3+B220+CD4−CD8− DNT cells are reduced in the lymph nodes of Sh3bp2KI/+Faslpr/lpr mice. (a,b) Lymph node cells were collected from WT (n = 7), Sh3bp2KI/+ (n = 7), Faslpr/lpr (n = 5), and Sh3bp2KI/+Faslpr/lpr mice (n = 6) at 48 weeks of age, and T cell subsets were stained with fluorochrome-labeled antibodies against CD3, CD4, CD8, CD25, and B220. (a) The ratio of T cell subsets was analyzed by flow cytometry. All cells were gated as 7-AAD-negative single cells, followed by being gated as lymphocytes. Values are presented as the mean ± SD; * P < 0.05; n.s. = not significant. (b) Representative flow cytometry plots of DNT cells in the lymph nodes. Flow cytometry shows a decreased proportion of DNT cells in the lymphocyte fraction of Sh3bp2KI/+Faslpr/lpr cells. The number in parentheses indicates the percentage of DNT cells in total lymphocytes. Note: DNT, double-negative T cell; SH3BP2, SH3 domain-binding protein 2; WT, wild-type; KI, knock-in; 7-AAD, 7-aminoactinomycin D.