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. 2021 Nov 26;10(12):3321. doi: 10.3390/cells10123321

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Purified EV identity. (A) Representative negative contrast electron microscopy of MOLM-14 EVs. (B) Representative cryo-electron microscopy showing EVs with typical double-membrane structures. (C) Mode size distribution using a 150 nm pore on a tunable resistive pulse sensing (TRPS) device (n = 3 measurements per cell line). (D) Representative super-resolution microscopy images (dSTORM mode, ONI) of MOLM-14 EVs stained with an anti-human CD63-AlexaFluor488 and an anti-human CD81-AlexFluor647 antibody. Scale bars represent 100 nm. (E) Western blots of identity markers according to the MISEV 2018 criteria suggesting absence of cellular contaminants in the EV purifications by lack of calnexin (90 kDa) and enrichment in small EV markers CD9 (25 kDa), CD63 (40–60 kDa), CD81 (26 kDa) and flotillin-1 (48 kDa). n = 3, representative blots shown.