Extracellular Vesicles from Caveolin-Enriched Microdomains Regulate Hyaluronan-Mediated Sustained Vascular Integrity

Figure 1

Determination of extracellular vesicle (EV) release kinetics and caveolin-enriched microdomain (CEM) dependence in human endothelial cells (EC). Panel (a): graphical representations of the kinetics of HA-induced EV release from human pulmonary microvascular endothelial cells (HPMVEC). Human EC monolayers were placed in serum-free media and the resulting media containing EV were collected at 0, 1, 2, 3, 6, 12, and 24 hours. The concentrations of EV were determined utilizing nanosight nanoparticle tracking analysis (NTA). HPMVEC exhibit basal secretion of EV (control). However, addition of 100 nM HMW-HA or 100 nM LMW-HA dramatically increased the production of EV starting at ~6 hours posttreatment. per group and error bars = standard deviation. Panel (b): graphical representation of the role of CEM in HA-mediated EV production from human EC. HPMVEC monolayers were either untreated or treated with 5 mM methyl-β-cyclodextrin (MβCD, a CEM disrupting agent) with or without addition of 100 nM LMW-HA or 100 nM HMW-HA (24 hours). EVs were analyzed as described in Panel (a). Inhibiting CEM formation significantly inhibited both LMW-HA and HMW-HA-mediated EV secretion with per group and error bars = standard deviation.

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