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. 2024 Dec 4;15:10543. doi: 10.1038/s41467-024-54850-6

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 1 — a Western blotting to validate EBV infection of GC tissues used for MV extraction. EBV⁺ and EBV⁻ human GC tumor samples are used for protein extraction and western blot. EBNA-1, Epstein-Barr virus (EBV)-encoded nuclear antigen 1, a marker protein of EBV. All lanes are loaded with 50 μg of total protein. b Centrifugation protocol and workflow for microvesicle enrichment from gastric cancer tissues. c Representative nanoparticle tracking analysis. d Transmission electron microscopy (TEM) imaging of MVs from gastric cancer tissues. Scale bar represents 100 nm. e Western blot of MVs isolated from gastric cancer for conventional extracellular vesicle markers. GAPDH glyceraldehyde-3-phosphate dehydrogenase. Western blotting of CD63 under non-denaturing conditions. The two lanes are loaded with the same volume (15 μl) of samples. f Volcano plots showing that among differential expressed proteins, 43 upregulated and 133 downregulated in MVs from EBV positive GC tissues relative to EBV negative tissues. g Heatmap showing the quantification of differentially expressed proteins. Upregulated proteins are listed in the enlarged panel. Results from 3 biological replicates. h KEGG pathway analysis for the 176-differential expressed MV proteins. Top 10 KEGG pathways are shown based on enrichment score. i Diagram shows the screening strategies for key MV proteins specific to EBV infection. Representative of two independent experiments (a, d, e).