fig6

Figure 6. Purification and analysis of exhaled extracellular vesicles from mouse EBC. EBC collected directly from the nose and mouth of individual animals was evaluated for the presence of exhaled EVS. (A) EBC from healthy control (n = 3) and lung tumor-bearing (n = 3) mice collected at weeks 2, 6, and 11 was evaluated using the Spectradyne nCS1 nanoparticle analyzer, using C400 cartridges for the detection of nanoparticles between 65 and 400 nm; (B) EBC samples from 6 control (left; 3 mL) and 3 lung tumor-bearing (right; 1 mL) mice, collected over 4 weeks, were subjected to ultracentrifugation and the pellets analyzed by imaging using transmission electron microscopy (TEM); (C) EV pellets from healthy control (top panels) and from lung tumor-bearing mice were analyzed using Super-Resolution Microscopy (ONi instrument) using anti-human anti-CD63, anti-CD9, and anti-CD81 anti-tetraspanin antibodies to evaluate the size and identity exhaled EVs contained in the ultracentrifuged EBC pellets of control (top panels) and lung tumor-bearing mice (bottom panels).