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[Preprint]. 2021 Dec 10:2021.12.04.471153. [Version 2] doi: 10.1101/2021.12.04.471153

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Figure 4. — Mechanically generated, cell-derived nanovesicles contain ACE2 and inhibit SARS-Cov-2 pseudotyped lentivirus transduction. A) Cartoon depicting the process used to isolate nanovesicles (NVs) used in this study. B) Representative histogram of nanoparticle tracking analysis of HEK293FT EV subpopulations as compared to NVs; data are normalized to the modal value in each population. C) Western blots of vesicles targeting standard EV markers CD9, CD81, and Alix. Western blot of vesicles and cell lysate of the contaminating endoplasmic reticulum protein, calnexin. D) Western blot against the c-terminal HA-tag of transgenic ACE2 in vesicle populations from parental or engineered cell lines, normalized by vesicle count. Results are representative of two independent experiments. E) Transmission electron microscopy images of NVs alongside micrographs of HS-EVs and UC-EVs. Scale bar represents 100 nm. F) Dose-response curve demonstrating the relationship between decoy vesicle dose and normalized percentage of cells transduced by Spike-lenti. Curves are normalized to the percent of cells transduced at the lowest vesicle dose in a particular curve. Symbols repesent the mean of three biological replicates; error bars are standard error of the mean. Data are representative of two independent experiments.