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Additional file 2: of Neutrophil elastase cleaves epithelial cadherin in acutely injured lung epithelium

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posted on 17.10.2016 by Rachel Boxio, Julien Wartelle, Béatrice Nawrocki-Raby, Brice Lagrange, Laurette Malleret, Timothee Hirche, Clifford Taggart, Yves Pacheco, Gilles Devouassoux, Abderrazzaq Bentaher
NE degrades cell-associated E-Cad. Confluent 16HBE cells were left untreated or treated with varying concentrations of purified NE (0, 2, 20, or 200 nM) for 6 h. Next, equal protein aliquots from cell lysates (10 μg) were subjected to SDS-PAGE and immunoblotting using antibodies raised against C-terminal parts of E-cad. A, Left panel. Anti-E-cad C-terminal antibody revealed a progressive decrease of E-cad that paralleled the increase of NE concentration. Right panel, densitometric analysis confirms decreased levels of E-cad. Data are mean values ± SD. *p < 0.05; Kruskall-Wallis test. Of note, anti-E-cad C-terminal antibody detected varying fragments. Immunoblotting for GAPDH, an internal control, was used as protein loading control of cell lysate proteins. Experiments were repeated three times. (TIFF 2703 kb)


Inserm Avenir Program, Agence Nationale de la Recherche, Fondation pour la Recherche Médicale, Fonds AGIR pour les Maladies Chroniques.