Th-POK inhibits the development of NKT17 cells in the thymus. (A) Plots depict NK1.1 and CD44 expression in Th-POK^hd/hd, Th-POK^hd/+, and Th-POK^+/+ littermate Vα14i NKT thymocytes, with gates shown used to define stage 1 (CD44^low, NK1.1⁻), 2 (CD44^high, NK1.1^−/low), and 3 (CD44^high, NK1.1^high) Vα14i NKT-cell subpopulations. (B) Bar graph shows stage 1-3 Vα14i NKT thymocyte numbers in sex- and age-matched Th-POK^hd/hd and Th-POK^hd/+ mice (n = 18). (C) Histogram overlays depict the expression RORγT, CD103, and CD196 in stage 2 Vα14i NKT cells from a set of Th-POK^+/+, Th-POK^hd/hd, and Th-POK^hd/+ littermates. Data are representative of 3 independent experiments. (D) Left scatter plot depicts the numbers of stage 2 NKT17 thymocytes from sex- and age-matched pairs of Th-POK^hd/hd and Th-POK^hd/+ mice (n = 7). Right scatter plot depicts the numbers of stage 2 NKT17 thymocytes from sex-matched littermates or age-matched pairs of Th-POK^hd/+ and Th-POK^+/+ mice (n = 6). P values were calculated using a paired Student t test of the log values. (E) Plot depicts IL-17 and CD103 staining of electronically gated stage 2 Vα14i NKT cells from Th-POK^hd/+ and Th-POK^hd/hd littermate thymocyte suspensions cultured with PMA and ionomycin. Data are representative of 4 Th-POK^hd/+ and 3 Th-POK^hd/hd samples examined in 2 separate experiments.