Immunohistochemistry of α-SMA, Col III, TGF-β1. Paraffin sections of liver were incubated overnight at 4°C with anti-α-SMA (Figure. 5A), anti-type III collagen (Figure. 5B), anti-TGF-β1 (Figure. 5C) monoclonal antibodies (1:100 dilutions). This was followed by incubation in biotinylated goat anti-mouse antibodies (1:200 dilutions) for 40 min and treated with SABC for 30 min at 37°C. The peroxidase activity, which resulted in a brown staining of cytoplasmic reaction sites, was visualized by incubation for 30 sec~1 min in dimethylbenzidine and staining lightly with hematoxylin.