Fig. 2. TGF-ß1-mediated EMT in NMuMG/E9 cells. (A) NMuMG/E9 cells were stained for E-cadherin (a,b), ZO-1 (c,d), paxillin (e,f) and F-actin (phalloidin; g,h) in the absence of TGF-ß1 (a,c,e,g) or in the presence of TGF-ß1 (5 ng ml^–1 for 1 day; b,d,f,h). In the presence of TGF-ß1, the signals for E-cadherin and ZO-1 became weak and punctate, and focal adhesions (f) and stress fibers (h) were induced. Photographs were taken using a 63x oil objective; scale bar, 10 µm. (B) NMuMG/E9 cells were treated with TGF-ß1 and cultured without passage for the indicated times. TNE extracts were immunoblotted for N-cadherin, E-cadherin, ß-catenin and GAPDH at the indicated times from t=0 hour (h) to t=9 days (d). Immunoblots were quantified and normalized to GAPDH as shown on the graph. ZO-1 (an epithelium marker) and fibronectin (a mesenchyme marker) were examined in the same time course.