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. 2023 Apr 12;14:2066. doi: 10.1038/s41467-023-37589-4

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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. 5 — a K19^Cre was induced in Krt19^CreERTtdTom embryos at E15.5 and after 24 h thymi were grafted under the kidney capsule of WT mice. Thymus grafts were harvested at the equivalent of postnatal week 4 and 8. b At 4 weeks, fate-mapped cells were detected within EpCAM1⁺ cells by flow cytometry and quantitated (n = 9, from 4 independent experiments). c Representative FACS plots and quantitation of K19-tdTom fate-mapped cells at 4 weeks within EpCAM1⁺UEA1⁺ and EpCAM1⁺Ly51⁺ TEC. d Representative FACS plots illustrating the phenotype of fate-mapped mTEC subsets within fate-mapped thymus grafts at postnatal week 4. mTEC^hi (MHCII^hiCD80^hi, n = 9), mTEC^lo (MHCII^loCD80^lo, n = 9), Aire⁺ (MHCII^hiCD80^hiAire⁺, n = 9), CCL21⁺ (n = 4) and tuft cells (MHCII^loCD80^loDCLK1⁺, n = 5), and corresponding quantitation. e Immunofluorescence of fate-mapped thymi at postnatal week 4, Aire (turquoise) K19-tdTom (red), K5 (blue). Scale bar denotes 10 μm. Image representative of 3 grafts. f At 8 weeks, fate-mapped cells were detected within EpCAM1⁺ cells by flow cytometry and quantitated (n = 10, from 4 independent experiments). g Representative FACS plots and quantitation of K19-tdTom fate-mapped cells at 8 weeks within EpCAM1⁺UEA1⁺ and EpCAM1⁺Ly51⁺ TEC. h Representative FACS plots illustrating the phenotype of fate-mapped mTEC subsets within fate-mapped thymus grafts at postnatal week 8. mTEC^hi (MHCII^hiCD80^hi, n = 8), mTEC^lo (MHCII^loCD80^lo, n = 8), Aire⁺ (MHCII^hiCD80^hiAire⁺, n = 8), CCL21⁺ (n = 5) and tuft cells (MHCII^loCD80^loDCLK1⁺, n = 4), and corresponding quantitation. i Immunofluorescence of fate-mapped thymi at postnatal week 8, Aire (turquoise) K19-tdTom (red), K5 (blue). Scale bar, 10 µm. Image representative of 3 grafts, from 3 independent experiments. j Representative FACS plots of SSEA1 and K19 expression at E12.5 ( j) and E15.5 (k), and quantitation of K19⁺ SSEA-1⁺ TEC (n = 12 for both stages, from 3 independent experiments). The data are shown as mean ± SEM.