Characterization ofex vivogenerated OT-I CD8 SP T cells. (A-B) Representative flow cytometric analysis of OT-I CD8^int (A) or CD8^hi (B) SP T cells phenotype on rIL-4 treatment. (C-D) Representative phenotypic analysis of OT-I CD8 SP T cells generated in the presence of rIL-7 (C) or rIL-13 (D). (E) Representative flow cytometric analysis of EOMES intracellular staining of OT-I CD8 SP T cells generated from the positive selection of DP3 thymocytes. The black line represents the isotype control and the red line represents EOMES staining. (F) qPCR analysis of Foxo1, Klf2, and S1pr1expression on ex vivo generated OT-I CD8 SP T cells. Data depict transcript abundance relative to untreated DP thymocytes. Three mice per group were tested. *P < .05; **P < .0004. For panels A through F, data shown are representative of 3 separate experiments.