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. 2019 Nov 21;10:2710. doi: 10.3389/fimmu.2019.02710

Figure 6.

Figure 6

Intrinsic control of iNKT cell effector lineage differentiation by Foxo1. (A–C) Thymocytes from mixed BM chimeric mice as described in Figure 2 were similarly analyzed. (A) Representative FACS plots showing RORγt and T-bet staining in gated CD45.1+ WT and CD45.2+ Foxo1KO iNKT cells (left panels) and PLZF and GATA3 staining in gated RORγt−T-bet− iNKT cells. (B) Scatter plots showing percentages of iNKT effector lineages. (C) Scatter plots showing CD45.2 Foxo1KO/CD45.1WT ratios of individual iNKT effector lineages. Data shown are representative or pooled from five experiments. Connecting lines indicate CD45.1 WT and CD45.2 Foxo1KO iNKT cells in individual mice. One chimeric mouse was examined in each experiment.*p < 0.05; **p < 0.01 determined by two-tail pair-wised Student t-test. (D–G) Two to three weeks old Foxo1f/f -Cd2iCre and Foxo1f/f mice were analyzed for thymic iNKT cells by flow cytometry. (D) Death rates of iNKT effector lineages detected by Live/Dead™ Fixable Violet Dead Cell Stain. N = 5. *p < 0.05 determined by two-tail pair-wised Student t-test. (E) Overlaid histograms show ROS levels in stages 1–3 iNKT cells. (F) Overlaid histograms show T-bet and CD122 levels in iNKT1 cells and RORγt levels in iNKT17 cells and in DP thymocytes. (G) Overlaid histograms show IL7Rα and ICOS levels in stages 1–3 iNKT cells. Data in (E–G) are representative of three experiments.