Fig. 2. Glycolysis is required for the development of effector CD8 T cells.

a Representative staining profile of CD25, CD44, and CD69 in WT and Pgam1 KO CD8 T cells stimulated with anti-TCR-β plus anti-CD28 mAbs for 36 h. b Representative staining profiles of phospho-STAT5 (Tyr694) in activated CD8 T cells. WT and Pgam1 KO naïve CD8 T cells were stimulated with anti-TCR-β mAb plus anti-CD28 mAb for 48 h, and then the cells were cultured with or without IL-2 for 24 h. c WT and Pgam1 KO naive CD8 T cells were labeled with eFluor670 and stimulated with anti-TCR-β mAb plus anti-CD28 mAb. The cell division was detected by flow cytometry at the indicated number of hours after the initial stimulation. d The immune response of antigen-specific CD8 T cells after Lm-OVA infection was analyzed by staining with an OVA-specific tetramer (Tet). A representative staining profile of Tet/CD8 gated on the CD8-positive cells in the spleen at 7 days post infection (upper). The percentages of cells are indicated in the circle. The absolute number of Tet⁺ CD8 T cells in the spleen is shown (lower). Each point represents an individual mouse. e Representative results of the intracellular FACS analysis of IFN-γ/IL-2 in the WT and Pgam1 KO CD8 T cells cultured under IL-2 conditions on day 5. The percentages of cells are indicated in each quadrant. f The results of an ELISA for IL-2, IFN-γ, and TNF-α in the supernatants of the cells in e (n = 3, biological replicate). The results of the analyses are representative of at least three independent experiments with similar results. The results are indicated with the standard deviation. *P < 0.05, **P < 0.01 (Student’s t-test).