Figure 1. CD1d-tetramer-based enrichment of thymic iNKT cells. (A) Identification of iNKT cells in BALB/c thymocytes by CD1d tetramer (CD1dTet) vs. TCRβ (top) or CD1dTet vs. CD24 analysis (bottom). Results are representative of 5 independent experiments. (B) MACS-based enrichment of CD1dTet⁺ iNKT cells is demonstrated by CD1dTet vs. TCRβ (top) or CD1dTet vs. CD24 analysis (bottom) of iNKT cells in total thymocytes or after MACS column enrichment. Results are representative of 5 independent experiments. (C) Percentages of iNKT cells in total thymocytes (before) and CD1dTet-enriched fraction (after). Plot shows summary of 5 independent experiments. (D) Surface TCRβ expression and CD1dTet staining on thymic iNKT cells before and after MACS-mediated enrichment for iNKT cells. Histograms (left) are representative and graphs (right) show summary of 5 independent experiments. (E) Intranuclear staining for PLZF and RORγt shows subset distribution before and after MACS-mediated enrichment for thymic iNKT cells. Enriched iNKT cells were stained for CD24 and gated on CD24^lo to identify mature iNKT cells. Dot plots (left) are representative and graphs (right) show summary of 5 independent experiments.

NS, not significant.

^**p<0.01; ^***p<0.001 were considered statistically significant.