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. 2001 Sep 17;194(6):797–808. doi: 10.1084/jem.194.6.797

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© 2001 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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BECs and LECs cannot be distinguished by conventional light microscopy when grown separate (A, BECs; B, LECs). (C) In mixed cultures, immunolabeling with anti-podoplanin (TRITC, red) and anti-vWF (FITC, green) reveal multicellular islands of podoplanin⁺/vWF⁺ LECs surrounded by podoplanin⁻/vWF⁺ BECs. BECs express more vWF than LECs (see also Fig. 3 A). Original magnification in A and B: ×300; C: 250. To confirm their phenotypic stability, cultured LECs and BECs were harvested, labeled with anti-podoplanin IgG, and analyzed by FACS ^® (insets in A–C; dotted line denotes the cut-off for podoplanin positivity). Cultured BECs (A) were homogeneously podoplanin-negative, cultured LECs (B) exclusively podoplanin-positive, while the mixed EC population (C) contained both qualities.