Figure 5 | Scientific Reports

Figure 5

From: Tumor-Derived Exosomes Induce the Formation of Neutrophil Extracellular Traps: Implications For The Establishment of Cancer-Associated Thrombosis

Figure 5

NETs interact with tumor-derived procoagulant exosomes. (A) Flow-cytometric analysis of TF expression in 4T1 cells. Black region represents labeling with a rabbit polyclonal anti-murine TF antibody and a phycoerythrin-conjugated secondary antibody. Gray regions represent cells labeled with IgG isotype control and the same phycoerythrin-conjugated secondary antibody. (B) Procoagulant activity of 4T1-derived exosomes. Exosomes were isolated and quantified from culture supernatants and further assayed for procoagulant activity, as described in the Methods section. Control bar represents the coagulation time of murine plasma alone. The asterisks indicate P < 0.001 relative to control plasma (Student’s t-test). Experiments were performed in triplicate. (C) Representative image showing 4T1-derived exosomes interacting with NETs. Exosomes were labeled with DilC18 (red), and NET DNA was stained with Hoechst 33342 (blue). Cells were isolated and stimulated with PMA for 3 hs to induce NET formation, before incubation with 4T1 exosomes. Scale bar = 20 μm.

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