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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. 5 — (a) Thymic B cells from wild-type, Tcra^–/–, and Cd40^–/– mice (all n = 5 biological replicates) were characterized for their expression of surface markers IgD, IgM, MHC class II, and CD80. Representative cytograms and histograms of thymic B cell subsets (DP = double positive IgM⁺IgD⁺, IgM⁺IgD^–, and DN = double negative IgM^–IgD^–) are shown along with mean fluorescence intensities (MFI) ± SD of MHC class II and CD80 next to the corresponding histograms. (b) Aqp4 expression in EpCAM⁺ thymic epithelial cells (TECs) isolated from wild-type and Cd40^–/– mice (n = 3 biological replicates). Mean RQ ± SD normalized to astrocytes. (c) Expression of Aqp4 in wild-type IgM⁺IgD^– thymic B cells vs. Cd40^–/– thymic B cells. Box plot derived from the RNAseq data in Fig. 3i, with the median as the centre, the first and third quartiles as the boundaries of the box, and 1.5 times the IQR as the whiskers. (d) Representative cytograms and quantification of P41/I-A^b-reactive T cells isolated from secondary lymphoid tissue (sec LyTi, spleen plus draining lymph nodes) of P41-immunized Aqp4^–/– and Cd40^–/– mice (both n = 3 biological replicates) on day 10 after immunization. Data are shown as mean ± SD tested with a two-tailed unpaired t-test, ns = not significant.

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