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. 2020 Jul 24;3:394. doi: 10.1038/s42003-020-01122-w

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a The intracellular amounts of ATP in CD8 T cells stimulated with anti-TCR-β mAb plus anti-CD28 mAb in the presence or absence of glutamine for the indicated number of hours. The results are presented with the standard deviations (n = 3, biological replicate). b The results of the immunoblot analysis of protein-tyrosine phosphorylation and β-actin in naïve CD8 T cells cultured with or without glutamine for the indicated number of hours. The protein amount of β-actin was used as a loading control. The numbers below the bands indicate the densitometry ratio with CD8 T cells cultured with glutamine. c The effect of rapamycin on glutamine-induced protein-tyrosine phosphorylation. Naïve CD8 T cells were stimulated under glutamine-sufficient conditions in the presence or absence of rapamycin for the indicated number of hours. The numbers below the bands indicate the densitometry ratio with CD8 T cells cultured without rapamycin. d The results of the immunoblot analysis of phospho-Zap70 (Tyr319), phosphor-Plcγ1 (Tyr783) and β-actin (control) in cells in (b). The numbers below the bands indicate the densitometry ratio with CD8 T cells cultured with glutamine. e The results of the immunoblot analysis of phospho-Zap70 (Tyr319), phosphor-Plcγ1 (Tyr783) and β-actin (control) in cells in (c). The numbers below the bands indicate the densitometry ratio with CD8 T cells cultured without rapamycin. f Representative results of the FACS analysis of CD25 and CD69 in CD8 T cells stimulated under the indicated conditions for 48 h. g Representative results of the intracellular FACS analysis of phosphor-Stat5 in CD8 T cells cultured with or without IL-2 stimulation. 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. The results of the FACS and immunoblot analyses are representative of at least three independent experiments with similar results. The results of the immunoblot analyses are presented as cropped images. The full-length blots are presented in Supplementary Fig. 18. *p < 0.05, **p < 0.01 (Student’s t test).