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. 2022 Jun 21;20:95. doi: 10.1186/s12964-022-00902-7

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — BGLF2-containing exosomes are released by infected cells. A Centrifugation protocol and workflow for separation and enrichment of exosomes and virions. B Exosome marker proteins were enriched in fractions #1-3 after iodixanol floating density gradient centrifugation. For immunoblotting, proteins in each fraction were concentrated by acetone precipitation. CD63 and CD81 served as exosome markers. The EBV genome in each fraction was quantified by qPCR. C and D Pool E (fractions #1-3) containing the BGLF2 protein. Each pool (E, fractions #1-3; V, fractions #5-6; and C, fractions #14-16) was analyzed by immunoblotting (C) and qPCR (D). Data are presented as the mean ± SE. Viral capsid antigen (VCA) p18 served as a virion marker. E Workflow for particle purification. F Akata(-) cells were incubated with purified particles for 24 h. GFP positivity was determined by FACS. The EBV genome of each pool was quantified by qPCR. The results are presented as the mean ± SE of four independent experiments after normalization to the EBV genome levels and as relative infectivity to pool V (infectivity value of 1). Double asterisks, p < 0.01