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. 2023 Oct 30;2(11):1015–1031. doi: 10.1038/s44161-023-00354-3

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© The Author(s) 2023

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. 3 — a, Schematic for mixed bone marrow chimera experiment. Ldlr^−/− mice were lethally irradiated and rescued by donor bone marrow from (50%) LysM^cre R26^tdTomato (WT) and (50%) Trem2^−/− mice. Recipient mice were rested for 8 weeks and then fed an HFD for an additional 8 weeks to induce atherosclerosis. b, Flow cytometry gating of blood immune cells (CD45⁺) after 8-week HFD feeding, showing ratio of monocytes derived from WT (tdTomato⁺) and Trem2^−/− progenitors. c, Confocal micrograph of whole-mount aorta showing tdTomato labeling (red) and CD45 (white) staining to define cellular contributions to foamy macrophages. Representative image from two independent experiments. d, Quantification of tdTomato⁺ cells in blood compared to foamy macrophages from whole-mount aorta images (n = 3 mice per group). Data are mean ± s.e.m. Student’s t-test, *P < 0.05. e, Foamy FACS was performed on CD64⁺CD11b⁺ macrophages isolated from mixed bone marrow chimera aorta. Macrophages were separated into tdTomato⁺ and tdTomato⁻ populations and then assessed for foamy representation by SSC and Bodipy (neutral lipid) staining. f, Flow cytometric overlap between tdTomato⁺ (red) and Trem2^−/− (blue) derived macrophages from digested atherosclerotic aorta. g, Quantification derived from flow cytometric foamy FACS comparing relative contribution to foamy macrophages (n = 4 mice per group). Data are mean ± s.e.m. Student’s t-test, **P < 0.01. KO, knockout. MACS, macrophage.

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