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. 2022 Feb 22;12:3027. doi: 10.1038/s41598-022-06809-0

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© The Author(s) 2022

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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Identification and characterization of circulating EVs in PSC patients. (A) Representative scanning electron microscopy images of circulating EVs isolated from PSC patients. (B) Dynamic light scattering analysis of the size³¹ of circulating PSC-derived circulating EVs (max peak: 396 nm). (C) Flow cytometry analysis of calcein/FITC + circulating EVs detected in healthy controls (n = 11) and PSC patients (n = 25). (D,E) Flow cytometry analysis of circulating EVs positive for hepatocyte markers SLC27A5 or ASGPR1. (F) Flow cytometry analysis of circulating EVs positive for cholangiocytes marker CK-19. (G) Western blot analysis of EV marker Alix and CD61 and cholangiocyte marker CK19 in healthy control and PSC-derived circulating EV protein lysates (samples derive from the same experiment and blots were processed in parallel). Values represent mean ± SD. Kruskal–Wallis test with post-hoc Mann–Whitney test and Bonferroni correction were used for statistical analysis.