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. 2024 Jan 23;25(3):552–561. doi: 10.1038/s41590-023-01738-0

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© The Author(s) 2024

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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Extended Data Fig. 9 — a, Relative abundance of mitochondrial M+1 malate in human CD8⁺ Teff cells treated with LA and/or LC and 2 mM ²H-LA for 2 hrs. n = 5. b, Quantification of MCT1/LDHb expression from western blotting in Fig. 5e. n = 3. c, Immunoblots of CTSD in human CD8⁺ Teff cells (left). Quantification of CTSD expression from western blotting (right). The control was set to 1. n = 3. d, CTSD activity was detected in human CD8⁺ Teff cells. n = 5. e, Relative abundance of intracellular DAG in human CD8⁺ Teff cells. n = 5. f, Immunoblots of ASM and PKCθ in the lysosome of human CD8⁺ Teff cells (left). Quantification of ASM/PKCθ expression from western blotting (right). The control was set to 1. n = 3. g, Immunofluorescence staining of PKCθ and Lamp2 in human CD8⁺ Teff cells. Scale bar, 5 μm (left). Pearson’s correlation coefficient between PKCθ and Lamp2 (right). n = 6 cells examined over 3 independent experiments. h, PKC activity of human CD8⁺ Teff cells was detected. n = 5. i, Lysosomal acidification of human CD8⁺ Teff cells was stained with LysoSensor. n = 8. j, Fluo-4 MFI of cytosolic calcium release in human CD8⁺ Teff cells. k, Immunoblots of TFEB in human CD8⁺ Teff cells treated with LA and/or LC (left). Quantification of TFEB expression in nucleus from western blotting (right). n = 3. l, Percentage of IL-2⁺, INF-γ⁺, CD107a⁺ cells in CD8⁺ T cells derived from human malignant pleural effusion was detected by flow cytometry. The box plots show the median and first and third quartiles and the whiskers show 1.5 × interquartile range on either side. n = 5 in LA_low and n = 7 in LA_high group. m, Schematic of experimental design for n. n, Percentage of transferred human CD8⁺ T cells was detected in spleen and blood from humanized NOG mice treated with or without LC. n = 5 mice. Data are mean±s.e.m. n = biological replicates unless stated otherwise. P values were calculated using unpaired two-tailed Student’s t test (l, n), one-way ANOVA for Dunnett’s multiple comparisons test (a-i and k), ns (not significant).

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