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. 2021 May 25;144(4):286–302. doi: 10.1161/CIRCULATIONAHA.120.052318

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© 2021 The Authors.

Circulation 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 Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

This article is made available via the PMC Open Access Subset for unrestricted re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the COVID-19 pandemic or until permissions are revoked in writing. Upon expiration of these permissions, PMC is granted a perpetual license to make this article available via PMC and Europe PMC, consistent with existing copyright protections.

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Figure 5. — Systemic–venous endothelial cells (ECs) localize to the bronchial vascular plexus and visceral pleura. Representative, serial immunofluorescent images of systemic–venous EC markers COL15A1 and VWA1, panvenous marker ACKR1 (all in red), panendothelial markers PECAM1 and CLDN5, and lymphatic marker PDPN (all in green) in (A) a bronchus with accompanying arteries and (B) visceral pleura. In (A), COL15A1, VWA1, and ACKR1 stain the small vessels of the bronchial vascular plexus, but not the accompanying arteries; PECAM1 and CLDN5 stain all vessels. In (B), COL15A1, VWA1, and ACKR1 stain the pleural vessels, but not the alveolar microvasculature; PECAM1 and CLDN5 stain all vessels. PDPN identifies bronchial (A) and pleural (B) lymphatic vessels. VWA1 exhibits off-target staining in smooth muscle cells, and ACKR1 in ciliated cells. Nuclei are counterstained throughout with DAPI (4′,6-diamidino-2-phenylindole). Conventional immunohistochemical images of shown markers can be found in Figure IV in the Data Supplement.