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. 2024 Feb 21;627(8003):407–415. doi: 10.1038/s41586-024-07079-8

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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/.

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Extended Data Fig. 7 — (a-c) Mixed bone marrow chimeras (MBMC) were generated by grafting congenically marked Rag1^–/– bone marrow engineered to retrogenically express AQP4-specific TCR clone 6 mixed with bone marrow from either wild-type or Aqp4^ΔB mice (4:1) into lethally irradiated Aqp4^–/– recipients and tested 6 weeks after engraftment. (a) Representative cytograms of the polyclonal and the retrogenic (AQP4-specific) thymic compartment facing a thymic environment equipped with either wild-type (n = 6 biological replicates) or AQP4 deficient B cells (n = 7 biological replicates). (b, c) CD5 and TCR-β expression in CD4⁺CD8⁺ double positive (DP) thymocytes. (b) Representative cytogram overlays and (c) quantification of DP thymocytes from the polyclonal and the retrogenic (AQP4-specific) thymic compartments. Symbols represent biological replicates. (d) MBMCs were generated as described in (a-c) with wild-type (n = 4 biological replicates) and Aire^ΔB (n = 3 biological replicates) donor mice (4:1). (e) Quantification of the thymic CD4⁺ single positive (SP) fraction. Data in (c, e) are shown as mean ± SD and tested with two-way ANOVA and Sidak’s post-test. *** P < 0.001, **** P < 0.0001. ns = not significant. The diagram in (d) was created using Servier Medical Art under a Creative Commons licence CC BY 3.0.

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