Fig. 1

Degradation of E-cad is NE concentration and incubation time dependent. Epithelial cell protein extracts containing E-cadherin (10 μg) were incubated alone or in the presence of varying concentrations of NE for various time periods. The reactions were resolved by SDS-PAGE under reducing conditions and visualized by immunoblotting. a-b, incubation with NE (0, 1, 5, 10, 25, 50 nM) for defined time point (30 min) resulted in an enzyme dose-dependent degradation of E-cad derived from 16HBE (a) or MLE15 (b) epithelial cell extracts. c, Incubation of MLE15 epithelial cell extracts with NE (50 nM) for defined time points (0, 10, 15, 20, 25, 30 min) resulted in stepwise degradation of E-cad. d, E-cad cleavage was largely abrogated in both 16HBE and MLE15 protein extracts when NE (50 nM) was pre-incubated with the physiologic serine proteinase inhibitor SLPI (100 nM). Molecular weight (MW) (kDa) standards are between panels. The findings are illustrative of at least three independent experiments