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. 2024 Dec 4;15:10566. doi: 10.1038/s41467-024-54780-3

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Flow cytometric plots of B1 cells in Nmur1^iCre-eGFP Id2^fl/fl (blue) and littermate Id2^fl/fl mice (white) in the peritoneal cavity in steady state. B1 cells were pre-gated on live CD45⁺ TCRβ^-. First gate shows B1 cells gated as IgM⁺ CD23⁻. Second gate shows B1 cells gated as CD19⁺ CD23⁻, further gating of B1 cells included CD11b. B1a and B1b cells were subdivided using the marker CD5. b Quantification of B1 cells as in (a) (Id2^fl/fl n = 4, Nmur1^iCre-eGFP Id2^fl/fl n = 3). c–f Flow cytometric plots and quantification of B1 cells in the (c), Thoracal lavage (Id2^fl/fl n = 6, Nmur1^iCre-eGFP Id2^fl/fl n = 4), d Omentum (Id2^fl/fl n = 4, Nmur1^iCre-eGFP Id2^fl/fl n = 3) e Lung (Id2^fl/fl n = 5, Nmur1^iCre-eGFP Id2^fl/fl n = 4) and (f) spleen (Id2^fl/fl n = 8, Nmur1^iCre-eGFP Id2^fl/fl n = 7). g Relative and absolute cell numbers of B1 cells across different ages in Nmur1^iCre-eGFP Id2^fl/fl (blue, 3 weeks n = 3, 13 weeks n = 4, 18 weeks n = 5, 30 weeks n = 4) and littermate Id2^fl/fl mice (black, 3 weeks n = 4, 13 weeks n = 4, 18 weeks n = 4, 30 weeks n = 5). h, i Flow cytometric plots of B1 cells in Nmur1^iCre-eGFP Gata3^fl/fl (green) and littermate Gata3^fl/fl (white) mice in the (h), peritoneal cavity (Gata3^fl/fl n = 4, Nmur1^iCre-eGFP Id2^fl/fl n = 5) and (i), thoracal lavage (Gata3^fl/fl n = 4, Nmur1^iCre-eGFP Id2^fl/fl n = 3) at steady state. j Histology section of omental Fat-Associated-Lymphoid-Clusters (FALCs) from the ILC2 reporter mouse Nmur1^iCre-eGFP Rosa26^flSTOP-RFP/+ (violet signal) stained with anti-KLRG1 (light-blue), anti-CD5 (dark blue), anti-B220 (red), anti-CD43 (green), and DAPI. Yellow arrows identify ILC2s (RFP⁺ KLRG1⁺ CD5⁻), blue arrows point towards B1 cells (B220^low, CD43⁺). Scale bar overview 100 µm, zoom 20 µm. Each symbol represents data from one mouse, mean +/− SD, all data are representative of at least two independent experiments. Statistical significance was determined by two-tailed unpaired Student’s t-test (b–i), *p < 0.05 **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data, including exact p-values, are provided as a Source data file Fig. 1.