Fig. 1.

Thymic iNKT subsets have different avidities for the PBS57–CD1d tetramer that correlate with expression of surrogate markers reflective of strength of signaling during selection. a Cells from the thymus of BALB/c IL-4 reporter KN2 mice were stained with anti-TCRβ mAbs and PBS57–CD1d tetramers. iNKT cells were then electronically placed on a grid consisting of 30 gates, allowing for the analysis of iNKT cells expressing different TCR levels and/or different binding to the PBS57–CD1d tetramer for a given level of TCR. b The proportion of iNKT1 (PLZF^lo, Rorγt⁻, Tbet⁺), iNKT17 (PLZF^int, Rorγt⁺, Tbet⁻), iNKT2 (PLZF^hi, Rorγt^–, Tbet^–, or hCD2⁺) as well as the proportion of CD44^hi cells in each gate was recorded and displayed as a heatmap. c Representative histograms for the expression of CD5, CD6, Ly6C, and Egr2 in each iNKT cell subset (as defined by the gating strategy shown on the left) is shown. Data are representative of n > 3 mice for each staining