Lymphoid organ cellularities and lymphocyte staining in WT, KI, and KO mice. (A) Total cellularity was counted in spleen and thymus from multiple experiments using age- and sex-matched littermates. The values shown are from both male and female mice between 8 and 16 weeks of age. Horizontal bars and numbers shown represent the mean value of each group. (B) Thymocytes from individual WT, KI, and KO mice were analyzed for expression of T cell (CD4⁺/CD8⁺) and NKT cell (NK1.1⁺ TCRβ⁺) markers by flow cytometry (n = 3/group; 8-12 weeks of age). The numbers shown represent the percentages of cells in each quadrant. The results shown are representative of 3 experiments. (C) Absolute numbers and percentage of thymic NKT cells from WT, KI, and KO mice. Means plus or minus SEM are shown. Statistical analysis was performed by one-way analysis of variance (ANOVA) using Prism. *P < .05 when compared with WT. (D) Splenocytes from WT, KI, and KO mice were analyzed for expression of NK cells (NK1.1⁺CD3⁻), NKT cells (NK1.1⁺CD3⁺), T cells (CD4⁺/CD8⁺) and memory CD8⁺ T cells (cells were gated on CD8⁺ CD3⁺ then gated on CD44^highIL-2Rβ⁺; n = 3/group; 8-12 weeks of age). Numbers shown represent the percentage of cells in each quadrant or gate. The results shown are representative of 3 experiments. (E) Absolute numbers and percentage of splenic NK, NKT, CD8⁺ cells, and memory CD8⁺ T cells from WT, KI, and KO mice. Means plus or minus SEM are shown. Statistical analysis was performed by one-way ANOVA using Prism. *P < .05 when compared with WT.