Fig. 2.

Aberrant expression of molecules affected by Eomes in iNKT terminal maturation and effector function. a Expression of PLZF, Rorγt, Gata3, and T-bet by thymic and splenic iNKT from WT and Eomes cKO mice detected by flow cytometry. b Quantitative PCR analysis of Eomes mRNA in thymic iNKT cells (TCRβ⁺CD1d-tetramer⁺) from WT mice: NKT0 (CD24⁺CD44⁻NK1.1⁻CD8⁻), NKT1 1(CD24⁻NK1.1⁺CD27⁺CCR6⁻), NKT2 (CD24⁻NK1.1^-CD27⁺CD4⁺), NKT17 (CD24⁻CD27⁻CD4−CCR6⁺CD103⁺). (n = 5, mean ± SEM) **p < 0.01, two-tailed Student’s t-test. c The percentage of NKT1 (PLZF^dimT-bet⁺), NKT2(PLZF^highGata3⁺), and NKT17(PLZF⁺Rorγt⁺) subsets in thymus and spleen of WT and Eomes cKO mice. (n = 5, mean ± SEM). *p < 0.05, **p < 0.01, Mann–Whitney. d Representative histograms showing expression of CD69, CD11a, NKG2d, CD43, NK1.1, CD122, and IL17Rb by thymic and splenic iNKT cells from WT and Eomes cKO mice (n = 5). Similar data were obtained from at least three independent experiments. e, f Thymic iNKT cells from WT and Eomes cKO mice were analyzed by RNA-Seq. Expression levels (expressed as FPKM) of selected NKT1 (e) or NKT2 (f) subset-specific genes of thymic iNKT cells were compared between WT and Eomes cKO mice (n = 3, mean ± SEM). (WT vs KO, *p < 0.05, **p < 0.01, ***< p < 0.001, two-tailed Student’s t-test)