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. 2008 Jul 28;182(2):315–325. doi: 10.1083/jcb.200710067

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© 2008 Niessen et al.

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.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Slug represses the endothelial phenotype and directly regulates the VE-cadherin promoter. (A) Analysis of endothelial marker expression by qRT-PCR in Slug-transduced HMEC (n = 3). *, P < 0.05. (B) Immunoblots for endothelial and mesenchymal markers in empty vector–, NotchICD-, and Slug-expressing HMEC. (C) EMSA using in vitro–translated luciferase (Luc) or Slug-FLAG protein and ³²P-labeled double-stranded oligonucleotides for an E-box cis element (−97) or two putative Slug E2-box motifs (−306 and −379) in the human VE-cadherin promoter. Supershift assays with anti–FLAG-M2 or IgG control antibodies and competition assays with 50× wild-type (wt) or mutant probes are also shown. (D) Promoter activity in endothelial cells cotransfected with vector or Slug plasmids and wild-type, E2-box mutant, or E-box mutant mouse VE-cadherin promoter-luciferase constructs (n = 4; each experiment performed in triplicate). *, P < 0.05. Error bars show SEM.