Fibrinolytic activity of exosomes and MVs from cancer cells on a fibrin clot. The analysis was performed on MDA-MB-231 cells (A-B), MCF7 cells (C-F), and NB4 cells (G-J) cell lines. Each experiment was repeated with 3 independent EV isolations. A fibrin clot was formed by the incubation of thrombin with pure fibrinogen. Exosomes and MVs (30 μg of protein) were added to the top of clot, followed by the addition of plasminogen (0.15 μM), with or without inhibitor (100 mM ε-AcA or 2.2 μM Apro). For a positive control, plasminogen (0.15 μM) and tPA (25 nM) were added to the fibrin clot. OD was measured at 350 nm every 20 minutes for 18 hours at 37°C. Results are represented as the mean ± SEM (n = 3 independent samples). The percentage of clot retention is shown for MVs (Ai) and exosomes (Bi) from MDA-MB-231 cells, MVs (Ci) and exosomes (Di) from MCF7 cells, MVs (Ei) and exosomes (Fi) from MCF7-PMA cells, MVs (Gi) and exosomes (Hi) from NB4 cells, and MVs (Ii) and exosomes (Ji) from NB4-ATRA cells. A representative photograph of the experiment is shown for MVs (Aii) and exosomes (Bii) from MDA-MB-231 cells, MVs (Cii) and exosomes (Dii) from MCF7 cells, MVs (Eii) and exosomes (Fii) from MCF7-PMA cells, MVs (Gii) and exosomes (Hii) from NB4 cells, and MVs (Iii) and exosomes (Jii) from NB4-ATRA cells. *P ≤ .05, **P ≤ .01, ****P ≤ .0001, Mann-Whitney U test (1 tailed).