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. 2020 May 14;10:7939. doi: 10.1038/s41598-020-64531-1

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© The Author(s) 2020

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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Infectious microvesicles (ImV) carry viral RNAs. (a) RT-qPCR data quantifying both (+) and (−) sense viral RNA (vRNA) from infectious microvesicles (ImV) and mock microvesicles (MmV) that were collected at 8 hpi (see Methods for details). vRNA was measured after extracellular vesicles underwent: 1) no treatment (labeled ImVs or MmVs), or 2) freeze-thaw & detergent (1% sodium deoxycholate) & RNase treatment to break open vesicles and degrade unencapsidated RNAs (labeled DRImV for infectious or DRMmV for mock-infected samples). RT-qPCR relative quantification was calculated as ΔC_t where ΔC_t = (C_t of endogenous control gene (GAPDH)) – (C_t of gene of interest (vRNA)), using GAPDH of whole cells for normalization. (b) Schematic of the experimental design for determining the infectivity of untreated infectious microvesicles (ImVs) as compared to infection by ImVs treated by either freeze/thaw alone (FT) or FT, detergent, and RNase (DR). Serial dilutions of sample were used in plaque assays to quantify the number of infectious sites by visual inspection for cell death.