Fig. 1.

Effects of ECAD on the expression of αSMA and vimentin. (A) ECAD loss in the fibrotic liver. Replicate liver sections prepared from rats that had been treated with vehicle (top) or DMN (10 μL/kg of body weight intraperitoneally 3 times per week for 4 weeks; bottom) were immunochemically stained for ECAD, αSMA, or GFAP (red). Arrowheads and arrows indicate the strong intensities of ECAD, αSMA, and GFAP. An arrow in the middle column shows αSMA expression around the vascular smooth muscle. The pictures are representative images from at least six independent experiments. The scale bar represents 20 (left and right) or 50 μm (middle). (B) Cadherin switching in HSCs. Primary HSCs were cultured in a growth medium for 6 or 12 days, and the cell lysates (30 μg each) were subjected to immunoblotting (left). The expression levels of ECAD and NCAD were also determined in the lysates of HepG2 cells, primary hepatocytes, or quiescent HSCs (epithelial type) and in the lysates of activated HSCs, LX-2 cells, or MEF cells (mesenchymal type; right). (C) Real-time PCR assays. The data are the means and standard errors of at least six separate experiments (significantly different versus day 0: **P < 0.01). (D) Effects of forced ECAD overexpression on αSMA and vimentin levels. HSCs and LX-2 cells were infected with adenoviral recombinant ECAD (multiplicity of infection = 50). Adenoviral GFP was used as a control. Results were confirmed by three separate experiments. (E) Real-time PCR assays in LX-2 cells infected with adenoviral recombinant ECAD (significantly different versus an adenoviral GFP infection: **P < 0.01 and *P < 0.05).