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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. 3 — a Flow cytometric plots and quantification of B1 cells in Il5^Cre/Cre (yellow, n = 4) and littermate Il5^Cre/+ mice (white, n = 7) in the peritoneal cavity and thoracal lavage at steady state. b Histology section of omental FALCs from Il5^Cre/+ Rosa26^flSTOP-YFP/+ mice. Staining with anti-B220 (red), anti-CD43 (green), anti-KLRG1 (light-blue), anti-CD5 (blue), endogenous IL-5 (Violet) and DAPI. Il-5 producing ILC2s (yellow arrows, Il5⁺KLRG1⁺CD5^-) reside in close proximity to B1 cells (blue arrows, B220^lowCD43⁺). Scale bars 50 µm. c Validation of the mouse line Nmur1^iCre-eGFP Il5^fl/fl by qPCR of sort-purified ILC2s from the lung (Il5^fl/fl n = 4, Nmur1^iCre-eGFP Il5^fl/fl n = 3). d IL-5 and Il-13 secretion was determined by flow cytometry of cultured immune cells isolated from the small intestine of Nmur1^iCre-eGFP Il5^fl/fl mice and littermate controls, stimulated with PMA/ionomycin (PMA/Iono) for 4 h (Il5^fl/fl n = 3, Nmur1^iCre-eGFP Il5^fl/+ n = 3, Nmur1^iCre-eGFP Il5^fl/fl n = 3). e, f Relative ILC2 numbers in the (e), small intestine and (f), mesenteric lymph nodes (MLN) (Il5^fl/fl n = 3, Nmur1^iCre-eGFP Il5^fl/fl n = 3). g–l Flow cytometric plots and quantification (Il5^fl/fl n = 3, Nmur1^iCre-eGFP Il5^fl/fl n = 3) of (g, i, k), B1 cells (cells were pre-gated on live CD45⁺ TCRβ⁻) and (h, j, l), CD11b⁺ B1 cells in the (g, h) peritoneal lavage, (i, j), the thoracal lavage and (k, l), the omentum. m Flow cytometric quantification of B1 cells in 2 weeks old Nmur1^iCre-eGFP Il5^fl/fl mice and littermate controls (Il5^fl/fl n = 9, Nmur1^iCre-eGFP Il5^fl/fl n = 8). n, o Flow cytometric plots and quantification of B1 cells in the (n), peritoneal cavity and the (o), thoracal lavage in littermate, co-housed Cd4^Cre Il5^fl/fl mice and controls (Il5^fl/fl n = 7, Cd4^Cre Il5^fl/fl n = 5). 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 (a, e–o) or one-way ANOVA (c, d), n.s. non-significant, *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. 3.