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. 2022 Mar 31;11(7):1184. doi: 10.3390/cells11071184

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© 2022 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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Characterization of ectosome samples isolated from conditioned media of normal thyroid follicular epithelial Nthy-ori 3-1 cells and thyroid anaplastic carcinoma 8305C cells. (A) Morphological characterization of Nthy-ori 3-1- and 8305C-derived ectosomes by transmission electron microscopy (TEM). Size distributions are presented on histograms. Mean diameter ± standard deviation was calculated for all observed vesicles (n) from a respective sample. (B) Nanoparticle tracking analysis (NTA) of Nthy-ori 3-1- and 8305C-derived ectosomes. Results from five independent measurements for each cell line are presented as graphs. The shaded area depicts standard deviation. (C) Representative Western blot of extracellular vesicle markers in whole-cell protein extracts (line C) and ectosome samples (line E). After being separated by 10% SDS–PAGE and transferred into PVDF membrane, 50 μg of proteins were probed with anti-HSP70 (1:2000), anti-CD63 (1:2000), and anti-ARF6 (1:500) as primary antibodies, and anti-mouse IgG-HRP (1:400) as a secondary antibody.