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. 2020 Jun 1;10:8852. doi: 10.1038/s41598-020-65873-6

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© The Author(s) 2020

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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Human OA subchondral bone has peripheral and cholinergic nerves. Immunofluorescence of peripherin and ChAT in human OA subchondral bone (see video 2) after the 3DISCO clearing protocol and analysis with Imaris. The colocalization of both markers showed that some subchondral bone peripheral nerves marked by Prph (white) are also cholinergic, as they showed ChAT immunofluorescence (red) as shown on the right while some did not expressed ChAT (left). ChAT: choline acetyltransferase, Prph: peripherin.