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. 2022 Aug 18;12:854126. doi: 10.3389/fcimb.2022.854126

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Copyright © 2022 Fitzpatrick, Nader, Watkin, McCoy, Curley and Kerrigan

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Isolation and characterisation of exosomes from healthy and infected endothelial cell supernatant (A). Immunoblot of exosomal markers Alix (45 kDa), CD63 (38 kDa), Tsg101 (47 kDa), and Flotiliin-1 (40 kDa) isolated from the serum of S. aureus infected endothelial cells and healthy control. Exosomes were isolated by differential centrifugation and nanofiltration. (B) Densitometry representing western blot data of exosomal markers. (C) Transmission electron microscopy images represent superparamagnetic capture beads incubated with healthy endothelial exosomes and S. aureus infected endothelial exosomes. Images were taken at 50,000X (top) and 150,000X (bottom) magnification. Scale bar: 1 µm. Arrows point towards CD63 bead alone and CD63 bead bound to exosomes. (D) Flow cytometry analysis of CD63 beads bound to either healthy or infected endothelial exosomes. Data is representative of 3 independent experiments.