Fig. 4: LTβR expression by thymic epithelium controls the intrathymic iNKT cell pool.

a Thymocytes from Foxn1^Cre and LTβR^TEC mice, analysed for expression of TCRβ and reactivity with the PBS57-loaded CD1d tetramer to identify iNKT cells. TCRβ⁺PBS57:CD1d⁺ cells were analysed for intracellular expression of PLZF, Tbet and RORγt to identify NKT1, NKT2 and NKT17 subsets as indicated. b Absolute numbers of iNKT cells and iNKT subsets in Foxn1^Cre controls (n = 11 biologically independent samples, closed symbols) and LTβR^TEC mice (n = 12 biologically independent samples, open symbols), over three independent experiments. Percentages relative to Foxn1^Cre controls are also shown. Significant P values using two-tailed unpaired t test as follows: no. of total iNKT P = 0.0002, no. of NKT1 P = 0.0003, no. of NKT2 P ≤ 0.0001, no. of NKT17 P = 0.0021, % Total iNKT P = 0.0001, % NKT1 P = 0.0003, % NKT2 P ≤ 0.0001, % NKT17 P = 0.0021. c Levels of expression of IL-17RB (IL-25R) on iNKT subsets; grey histograms represent isotype controls. Bar graphs show MFI of IL-17RB on indicated iNKT subsets, n = 5 biologically independent samples, over three independent experiments. Significant P values using two-tailed unpaired t test as follows: NKT1 vs. NKT2 P ≤ 0.0001, NKT1 vs. NKT17 P = 0.0005. d Levels of CD122 expression on iNKT cell subsets; grey histograms represent isotype controls. Bar graphs show MFI of CD122 on indicated iNKT subsets, n = 5 biologically independent samples, over three independent experiments. Significant P values using two-tailed unpaired t test as follows: NKT1 vs. NKT2 P ≤ 0.0001, NKT1 vs. NKT17 P = 0.0004. e qPCR analysis of Il25, Il15 and Il15ra mRNA in CD104⁺ (closed symbols) and CD104⁻ (open symbols) mTEC^low isolated from adult WT mice. f qPCR analysis of Il25 in CD104⁻ mTEC^low, and expression of Il15 and IL15ra in CD104⁺ mTEC^low from control Foxn1^Cre (closed symbols) and LTβR^TEC mice (open symbols). All data are represented as mean ± SEM, **P < 0.01, ***P < 0.001 and ****P < 0.0001. qPCR was performed in replicate, and data shown are representative of three independent biological sorted samples. Source data are provided as a Source Data file.