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. 2021 Feb 3;9(2):146. doi: 10.3390/biomedicines9020146

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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 (http://creativecommons.org/licenses/by/4.0/).

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Characterization of EVs isolated from the CM of GSCs by transmission electron microscopy (TEM) and Western blot analysis. (a) Representative TEM images of two different types of EVs isolated from the total secretome of human GSCs. Left panel: MVs (Fn1 fraction) in the size range of 100–1000 nm and above. Right panel: Exo-like vesicles (some of which are indicated by black arrows, Fn5 fraction) in the size range of 30–100 nm. (b) Western Blot analysis of 30 µg of proteins from isolated EVs confirmed Fn5purity for the presence of canonical exosome proteins like Alix and CD63 and the absence of Calnexin, detectable only in the whole cell lysate (CL). Fn1 strongly reacted with anti-Abs to EpCAM, but no response was visible as for Alix or CD63. Mr = molecular range of the weight of proteins, expressed as kiloDaltons (kDa), revealed by the appropriate antibodies (see Methods Section).