E cadherin

For immunohistochemistry and Western blot analysis, β-actin (Sigma Inc.), α-smooth muscle actin (α-SMA, Sigma Inc.), E-cadherin (BD Transduction Lab., San Diego, CA, USA), and F4/80 (Serotec Inc., Raleigh, NC, USA) were used.
The wax of the 4 μm sectioned tissues was removed through alcohol and xylene, and the sections were then immersed in 0.4% methanolic H₂O₂ for 30 minutes, followed by 0.05% Triton X for 15 minutes. After incubation in 2% donkey serum (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) for 1 hour, these sections were immersed in primary antibodies, α-SMA 1:1,000, E-cadherin 1:200, and F4/80 1:50 overnight at 4ºC.
The following day, the sections were washed with 0.01 M PBS and then immersed in donkey-anti-rat or donkey-anti-mouse (Jackson ImmunoResearch Laboratories) secondary antibodies diluted to 1:200 and incubated for 2 hours at room temperature. Immunoreactivity was detected by incubation in Tris buffer (pH 7.6) containing 0.05% 3,3′-diaminobenzidine-4HCl and 0.005% H₂O₂. These sections were then counterstained with Meyer’s hematoxylin and mounted with Canada balsam for examination.