Jα18−/− mice were lethally irradiated, reconstituted with 95% Jα18−/− bone marrow and 5% bone marrow of the indicated TN lines, and analyzed 8 weeks later. (A) Thymocytes from C57BL/6 and all TN lines were stained with anti-CD3 and CD1d (PBS57) tetramer, and analyzed by flow cytometry. Mean fluorescence intensities (MFI) of tetramer staining for gated populations are shown on plots. (B) Thymocytes from C57BL/6 and all TN lines were stained with anti-CD3 and CD1d (PBS57) tetramer before being fixed, permeabilized, and stained with antibodies to PLZF, T-bet, and RORγt. Results shown are gated on CD1d (PBS57) tetramer+ iNKT cells. Tetramer bright = top 20% of iNKT cells; tetramer dim = bottom 20% of iNKT cells as defined by intensity of PE signal. (C) Quantification of mice shown in panel A. (D) Quantification of mice shown in panel B. n=4 mice per group. Error bars are SD.