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. 2022 Sep 16;7:53. doi: 10.1038/s41536-022-00250-7

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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. 3 — A Heatmap of clustering dysregulated mRNA expression profiles with microarray in T3-EV-treated BMSCs compared to un-EV treated control. B Volcano plot of mRNA expression profiles in T3-EV-treated BMSC recipient. C Dysregulated typical chondrogenic markers derived from the microarray results with T3-EV treatment. D All differentially expressed genes were subjected to gene ontology (GO) analysis (DEG with fold change >2 or <0.5, p value <0.01). BP biological processes, MF molecular function, CC cellular component. E Significantly enriched pathways for dysregulated genes enriched with T3-EV treatment in KEGG pathways. F, G FOXO1 expression(red) assayed with western blotting and fluorescent immunostaining (red for FOXO1; green for cytoskeleton) in T3-EV treated BMSC recipient. Cells were counterstained with DAPI for the nucleus (blue). Treatment with saline served as control. H Quantification of gene expression with the same BMSCs and the same EVs (n = 3 for each) with qRT-PCR for Chondrogenic genes (SOX9, ACAN, COL2A1, and MMP13) and FOXO signaling-related genes (FOXO1, Gadd45a, p27, and Cathepsin L). *P < 0.05, **P < 0.01, ***P < 0.001, NS not significant.