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. 2022 Aug 5;13(8):681. doi: 10.1038/s41419-022-05089-w

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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 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. 2 — a Cx43 levels detected by WB in sEVs released by OA-derived chondrocytes treated with oligomycin (Oligo). n = 3, Student’s t test. b Cx43-positive sEVs increased Cx43 levels in target chondrocytes treated for 48 h. Oligomycin (Oligo) treatment (5 µM, 48 h) was used as positive control for Cx43 overexpression. n = 4. c WB and flow cytometry analysis shows increased levels of the mesenchymal markers CD105 and CD166 in OA-derived chondrocytes after 48-h treatment with sEVs derived from Oligo-treated chondrocytes. n = 3, Student’s t test. d Immunofluorescence for collagen type II (Col2A1) showed a significant decreased in OA-derived chondrocytes treated with hCx43-sEVs for 48 h (n = 4, one-way ANOVA). On the right, RNA expression of the main cartilage ECM markers ACAN and Col2A1 also showed a significant downregulation after a 48-h treatment with hCx43-sEVs (n = 3, Student’s t test). e IL-1β overexpression in OA-derived chondrocytes treated with sEVs derived from Oligo-treated chondrocytes. n = 3, Student’s t test. f MAPK/ERK pathway is activated in OA-derived chondrocytes after hCx43-sEVs treatment, as detected by higher ERK1/2 levels detected by WB. n = 3, one-way ANOVA. g Correlated to gene expression, protein levels of the EMT-transcription factors Smad2/3 (n = 4) and Twist-1 (n = 3) were increased in OA-derived chondrocytes after a 48-h treatment with hCx43-sEVs. Student’s t test. h The treatment with Cx43-positive sEVs was correlated with overexpression of several EMT-related transcription factors. n = 3, Student’s t test. Data are expressed as mean ± S.E.M., *P < 0.05, **P < 0.01, ***P < 0.0001.