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Size and concentration measurements and transmission electron microscopy (TEM)
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For size and concentration measurements, 5–10 μg of EVs were resuspended in 1 mL of PBS and measured with a NanoSight NS300 instrument (Malvern, UK).
For TEM, 10 µL (100 µg/mL) of purified EVs were placed on non-glow-discharged carbon-coated Formvar/Carbon grids (PolySciences) and negatively stained for 1 min with 10 µL of 2% osmium tetroxide solution (Sigma-Aldrich). Negative stain solution was removed by wicking onto filter paper, and TEM images were obtained using a JEM-1400 Flash transmission electron microscope (Jeol, Japan).
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