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Fig. 5. Cadherin switching was reversible in TGF-ß1-treated NMuMG/E9 cells. (A) Untreated NMuMG/E9 cells (a,d,g), NMuMG/E9 cells treated with 1 ng ml–1 TGF-ß1 and 20 ng ml–1 EGF for 30 days (b,e,h), and NMuMG/E9 cells treated with TGF-ß1 and EGF for 23 days and subsequently cultured upon removal of both reagents for an additional 7 days (c,f,i) were stained for E-cadherin (a-c), ZO-1 (d-f) or F-actin (phalloidin) (g-i). Photographs were taken using a 40x dry objective; scale bar, 10 µm. (B) TNE extracts from untreated NMuMG/E9 cells (lane 1), TGF-ß1- and EGF-treated NMuMG/E9 cells for 60 days (lane 2) or reversibly treated NMuMG/E9 cells (lane 3) were examined for N-cadherin, E-cadherin, fibronectin and GAPDH by immunoblot. (C) RT-PCR was performed using total RNA extracted from untreated NMuMG/E9 cells (lane 1), TGF-ß1- and EGF-treated NMuMG/E9 cells (for 1 day, lane 2, or for 30 days, lane 3) or reversibly treated NMuMG/E9 cells (lane 4) to examine the levels of mRNAs encoding E-cadherin, N-cadherin, snail, SIP1 and GAPDH.





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