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. 2021 Jul 15;41(9):2454–2468. doi: 10.1161/ATVBAHA.121.316539

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© 2021 The Authors.

Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited.

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Figure 2. — Cell-type specific extracellular vesicle (EV) labeling strategy and live imaging in the pericardial space (PS).A, Ventral view of 3 dpf Tg(myl7:HRAS-mCherry) zebrafish, boxed region highlights position of B and C. B and C, Overview images of the entire hearts of Tg(myl7:HRAS-mCherry) fish in ventral view. B, A maximum projection of a fixed fish; (C) a single plane of live light sheet imaging. D, Image sequences of higher magnification views of the color coded boxed areas in C. mCherry+ CM-EVs are observed moving through the pericardial space (arrowed and inset). E–G, A maximum intensity projection of deconvolved images of the ventricle and internal surface of the pericardial wall (E) reveals static CM-EVs as shown with digital zoom of boxed region (F). The orthogonal view (YZ) of this region suggests the EVs may be associated with a layer of unmarked ECM (extracellular matrix) rather than direct contact with underlying cells (G). The magenta dashed line in B and C demarks the pericardial wall. The white dashed line in D demarks the ventricle (V) and atrium (A). Arrows indicate static EVs. Anterior is to the left. Scale bars: A, 100 μm; B and C, 50 μm; D, 20 μm; insets in D, 2 μm; E and G, 5 μm; F, 2 μm.