Fig. 3. E-Cadherin continued to function after morphological EMT was induced. (A) Permeabilized (0.2% Triton X-100, Tx100; a) and unpermeabilized (b) NMuMG/E9 cells that had been treated with 5 ng ml^–1 TGF-ß1 for 1 day were stained with antibodies against the extracellular portion of E-cadherin (ECCD2). E-Cadherin was localized to the cell surface (b). Photographs were taken using a 40x dry objective; scale bar, 10 µm. (B) Extracts of untreated NMuMG/E9 cells (lanes 3, 4) or NMuMG/E9 cells treated with 5 ng ml^–1 TGF-ß1 for 1 day (lanes 5, 6) were immunoprecipitated (IP) with anti-E-cadherin (EC; lanes 4, 6) or control antibodies (Ctr; lanes 3, 5) and immunoblotted for E-cadherin, ß-catenin and -catenin. Comparable amounts of ß-catenin and -catenin were coimmunoprecipitated with E-cadherin in treated (lane 6) and untreated (lane 4) NMuMG/E9 cells, indicating that E-cadherin junctional complexes were functional. Lanes 1, 2 are immunoblots of treated and untreated cells to show N-cadherin and fibronectin were induced in this experiment, as expected, and to show the input levels of E-cadherin, ß-catenin and -catenin in the immunoprecipitations.