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. 2023 Aug 9;9:290. doi: 10.1038/s41420-023-01588-7

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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. 3 — A Methodological approach for whole brain processing with CLARITY after the administration of IgG on the olfactory mucosa (cartoon produced with the Biorender software). B Representative images of IgG immunostaining on sagittal optical sections of mouse brains acquired by light-sheet microscopy, after tissue clearing and IgG detection by immunofluorescence. Scale bar = 1 mm. C Two different thresholds were applied to discriminate IgGs accumulated in the neuropil (low fluorescence intensity) and the vasculature (high fluorescence intensity). Scale bar = 500 μm. D Neuropil profiles of IgG fluorescence between the olfactory bulb and the cerebellum (1 brain/condition). E Average IgG fluorescence in blood vessels (1 brain/condition). Student’s t-test, ***P < 0.001. F, G Average fluorescence intensity of the neuropil (F) and vasculature (G) of brains undergoing different treatments. Each dot corresponds to the mean fluorescence intensity of a brain from independent experiments, normalized to the highest mean value for each set of experiments. n = 3 brains for EAE + 11C7, 3 for EAE + Ctrl IgG, 2 for EAE + vehicle, 1 for 11C7.