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

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)