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. 2024 Oct 2;16(11):2946–2975. doi: 10.1038/s44321-024-00145-8

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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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) UMAP projection of 194,999 randomly selected sub-sampled cells from E16.5 and P3 liver and P3 spleens, analyzed by 25-color flow cytometry, with cell type-annotated clusters, matched to (Fig. 2A) insofar as possible. (B) Heatmap showing a row z-score of median protein expression levels of cell type markers in the aggregated 25-color flow cytometry dataset from E16.5 and P3 livers, and P3 spleens of Jag1^+/+ (n = 7), Jag1^Ndr/+ (n = 11), and Jag1^Ndr/Ndr (n = 6) animals. (C, D) Representative flow cytometry plots (C) and relative frequency of the CD3⁺, CD4⁺/CD3⁺, and CD8⁺/CD3⁺ T cells in livers and spleens from the Jag1^+/+, Jag1^+/Ndr and Jag1^Ndr/Ndr mice at P3 (D). (E, F) Representative immunofluorescent staining (E) and quantification (F) of LCK⁺ T cells in periportal areas in Jag1^Ndr/Ndr and Jag1^+/+ livers at P10 (n = 5 Jag1^+/+, and 6 Jag^Ndr/Ndr animals). PV, portal vein. Scalebar lengths in (E) are specified within the panel and are identical for Jag1^+/+ and Jag1^Ndr/Ndr mice. HSc hematopoietic stem cells, EMP erythroid-myeloid progenitors, MPO myeloid progenitors, NK natural killer. All graphs represent mean ± SD. Statistical test in (D) is one-way ANOVA with Dunnett’s multiple comparison test, (Jag1^+/+ vs. Jag1^Ndr/Ndr) Jag1^+/Ndr and Jag1^Ndr/Ndr, and in (F) is an unpaired, two-tailed Student’s t-test, *p < 0.01); Each data point represents a biological replicate.