Figure 5.

SAP-dependent PLZF⁺ T cells were required for the innate-like CD8 phenotype in Id3^{−/ −} mice. (A) WT and Id3^{−/ −} thymocytes were examined by flow cytometry for intracellular PLZF (upper, grey histogram is isotype control) and PLZF⁺ cells were analyzed for TCRβ and TCRγ δ . PLZF⁺TCRβ ⁺ cells were analyzed for CD4 and CD8. PLZF⁺TCRβ ⁺CD4⁺ cells analyzed for staining with CD1d tetramer loaded with PBS57 (lower, grey histogram is unloaded tetramer). (B) Average number (+ standard deviation) of PLZF⁺TCRβ ⁺ cells in the thymus of WT and Id3^{−/ −} mice. n > 5 for each genotype. (C) CD4 thymocytes were analyzed for intracellular expression of IL4 and IFNγ 5 hours after stimulation with PMA + ionomycin. (D) Intracellular staining for PLZF (black line) in mice of the indicated genotype. Isotype control (grey). (E) WT, Zbtb16^lu/lu, Id3^{−/ −} and Id3^{−/ −}Zbtb16^lu/lu thymocytes were analyzed by flow cytometry for TCRβ (upper) and TCRβ ⁺ cells were analyzed for CD4 and CD8 (lower). The frequency of TCRβ + cells and CD4 or CD8 cells among TCRβ ⁺ cells is indicated. (E) Expression of CD122, CD44 or CD24 on TCRβ ⁺CD8 thymocytes from WT (grey) or mutant mice (black). Data is representative of 3 experiment.