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. 2022 Jun 29;5:633. doi: 10.1038/s42003-022-03569-5

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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. 7 — a Representative, fluorescence calibrated data obtained for buffer control (fPBS, left column) and PPP (right column) samples stained with anti-CD31-BV421 and anti-CD9-APC mAbs. Detergent treatment was performed by incubating the samples for 30 minutes with 20 µL 10% (v/v) TritonX-100 stock solution. Red gate: Single-positive CD9 events, purple gate: single-positive CD31 events, tan gate: double-positive events. I, double staining and II, double staining after detergent treatment. b Visual interrogation of the gated populations in the representative PPP sample. c Quantification of double-positive fluorescent events in 5 PPP samples and fPBS, stained with mAbs or isotypes, before and after detergent treatment. Approximately 93% of double-positive events in PPP stained with mAbs represent PPP-derived single EVs that were detected well above the fluorescent background. Red dots: means of sample spread. Symbols: individual PPP samples.