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. 2023 Nov 8;623(7988):803–813. doi: 10.1038/s41586-023-06717-x

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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 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. 5 — a, Immunofluorescence staining of thymic tissue from age-matched controls, a patient with autosomal-recessive complete RELB deficiency or heterozygous for a p52^LOF/IκBδ^GOF NF-κB2 variant. AIRE-expressing cells (green) and Hassall’s corpuscles (HaC) are shown on the left. Pan-K, pan-keratin. Staining for K10 (red), defining terminally differentiated corneocyte-like mTECs, is shown on the right. DAPI staining is shown in blue. Scale bars, 50 μm (left) and 100 μm (right). Inset: the controls at a higher magnification. Data shown are representative of one independent experiment. b, The percentage of EPCAM⁺CD45⁻ thymic epithelial cells (TECs), and the various TEC subsets (defined on the basis of their MHC class II (MHC-II) and AIRE expression) in WT controls (Nfkb2^+/+, black dots, n = 5) and mice carrying a heterozygous missense variant homologous to the human Y868* p52^LOF/IκBδ^GOF NF-κB2 variant (Nfkb2^+/Y868*, red dots, n = 7). Statistical comparisons were performed using unpaired, parametric, two-tailed Student’s t-tests (EPCAM⁺ TECs) or two-way nonparametric analysis of variance (ANOVA) (Sidak’s test) with correction for multiple comparisons (TEC subsets). Data are mean ± s.d. Data shown are representative of three independent experiments. c, Representative confocal microscopy images of AIRE (green), K5 (red) and DAPI (blue) of WT (Nfkb2^+/+, n = 3, top) and Nfkb2^+/Y868* (n = 3, bottom) mouse thymuses. Scale bars, 20 μm. Data shown are representative of two independent experiments. d, Mean fluorescence intensity (MFI) of AIRE expression in mature MHC-II^highAIRE⁺ mTECs from WT (n = 5) and Nfkb2^+/Y868* (n = 7) mouse thymuses. Statistical comparisons were performed using unpaired, parametric two-tailed Student’s t-tests. Data are mean ± s.d. Data shown are representative of three independent experiments.

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