RORγt⁺ iNKT cells respond weakly to IL-15 but are highly sensitive to TGFβ. Flow cytometry analysis to compare IL-15Rβ (CD122) and TGFβRII surface expression, and intracellular phospho-STAT5 (p-STAT5) and phospho-SMAD2/3 (p-SMAD2/3) levels in RORγt⁺ iNKT, RORγt⁻ iNKT, and non-iNKT cells in the thymus of 6- to 8-week-old C57BL/6 WT mice. Dashed lines represent staining of the secondary antibody without the primary antibody. For p-STAT5 staining, cells were incubated with IL-15. Invariant NKT cells were identified by excluding B220⁺ and CD8⁺ cells and gating on CD1d tetramer⁺ TCRβ⁺ cells. Data are representative of at least 3 experiments with 1 to 2 individual mice per experiment.