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. 2020 Jan 23;106(2):454–463. doi: 10.3324/haematol.2019.220210

Figure 5.

Figure 5.

Influence of polarization conditions on extracellular vesicle production of human macrophages. (A) Numbers of extracellular vesicles in the supernatant of unpolarized macrophages and macrophages polarized in the presence of 100 ng/mL lipopolysaccharide (LPS) and 100 ng/mL interferon (IFN)-γ or 20 ng/mL interleukin (IL)-4 and 20 ng/mL IL-13 were determined by flow cytometry as indicated in the Methods section. Values are given as total vesicle count per mL: n=9 for 4 h and 12 h; n=12 for 24 h; and n=14 for 48 h. (B) Phosphatidylserine (PS) content of extracellular vesicles in the supernatant of M0 and macrophages polarized in the presence of 100 ng/mL LPS and 100 ng/mL IFN-γ or 20 ng/mL IL-4 and 20 ng/mL IL-13 for 48 h was determined using a specific enzyme-linked immunosorbent assay as indicated in the Methods. Values are given in nM PS (n=6). (C) Total extracellular vesicle-derived RNA was evaluated in the supernatant of M0 and macrophages polarized in the presence of 100 ng/mL LPS and 100 ng/mL IFN-γ or 20 ng/mL IL-4 and 20 ng/mL IL-13 for 48 h as indicated in the Methods section. Values are given in ng/mL RNA (n=5) and represent mean values ± standard deviation. M0: unpolarized macrophages; M(LPS+IFN): classically activated polarized macrophages; M(IL-4+IL13): alternatively activated polarized macrophages.