Phenotype and expression of transcription factors in iNKT cells from Runx3-deficient, ThPok-deficient, DKO and their littermate controls. (a) Liver MNCs were purified from the indicated mice, gated on αGalCer+TCRβ+ iNKT cells, and further assessed for CD69 and CD62L expression by flow cytometric analysis. (b) Bar graphs show the MFI of CD69 and CD62L in the indicated iNKT cell populations from liver MNCs. Data are representative of four mice analyzed in five independent experiments. Bars depict the mean value of all samples collected. *P<0.05, **P<0.01, ***P<0.001. (c) Thymic iNKT cells were sorted to detect the expression of T-bet, GATA-3 and RORγt by RT-PCR. Data are representative of five independent experiments. *P<0.05, **P<0.01, ***P<0.001. DKO, double knockout; iNKT, invariant natural killer T; MNC, mononuclear cell; RORγt, receptor-related orphan receptor-γt; Runx3, runt-related transcription factor 3.