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Figure 2. EVs isolated from the blood plasma of healthy volunteers. (A) Size and concentration of non-permeabilized and permeabilized EVs purified from peripheral blood were determined by NTA; (B and C) Plasma EVs were characterized by immunoblotting (B) for the indicated proteins and dSTORM imaging (C) for CD9 (blue), CD63 (yellow), and CD81 (red). A magnified view of a single EV is presented (C); (D) Non-permeabilized and permeabilized EVs were immunolabeled for Alix or Syntenin (blue), CD63 (yellow), and CD81 (red) and analyzed by dSTORM; (E and F) EVs expressing single-, double-, and triple-positive of specific protein markers as indicated were quantified. For all EV quantification, a range of 2,000 to 3,000 EV particles were analyzed per replicate per condition. Scale bars, 10 µm (C, overview) or 100 nm (C, magnified, D).