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. 2021 Nov 3;599(7885):471–476. doi: 10.1038/s41586-021-04082-1

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© The Author(s) 2021

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. 4 — a, Quantification of mouse bone marrow or human monocytes cultured in the presence of macrophage colony-stimulating factor (M-CSF) for 6 d (mouse, n = 6) or 7 d (human, n = 5) with (GABA M-0) or without (M-0) 1 mM GABA. b, Flow cytometry of GABA M-0 and M-0 cell viability (assessed with propidium iodide (PI) and Annexin V, n = 4), F4/80, CD68 and FRβ (mouse, n = 3; human, n = 5)). c, Pathway analysis of mouse GABA M-0 and M-0 transcriptomes (n = 2) and proteomes (n = 3) (red, upregulated (>0); purple, downregulated (<0)). d, Gene expression of mouse M-0 and GABA M-0 cells after 6 h of treatment with IL-10 (M-IL-10 and GABA M-IL-10) (n = 3). e, Day 7 MC38 tumour monocytes differentiated in vitro into tumour monocyte-derived macrophages (TMDMs) with or without treatment with 1 mM GABA. Immunocytochemistry shows nuclear (TO-PRO-3) localization of total NF-κB p65 (t-p65) after TNF or IL-1β stimulation. Scale bars, 100 µm (n = 60). AU, arbitrary units. f, MC38 tumour volume in Mb1^cre/+;Gad1^fl/+ (n = 5) or Mb1^cre/+;Gad1^fl/fl (n = 6) mice. g, Flow cytometry of intracellular cytokines in CD8⁺ tumour-infiltrating lymphocytes (TILs) after re-stimulation (Mb1^cre/+;Gad1^fl/+, n = 4; Mb1^cre/+;Gad1^fl/fl, n = 5). Significance was calculated by two-tailed unpaired t-test (a, d) or two-way ANOVA (e, f): *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; NS, not significant. Data are shown as mean ± s.e.m. n indicates the number of biological (c, d, f, g) or technical (a, b, e) replicates. Data are representative of two (c (proteome)) or three (a (mouse), b (mouse)) experiments. Exact P values are provided in the source data.

Source data