Western blot analysis of xenograft tumors

Protein was extracted from resected tumors and western blot analysis was performed as previously described [39]. Briefly, for protein extraction, tumor samples were minced and incubated in lysis buffer (250 mmol/L Tris-HCl (pH 6.8), 2% SDS, and 10% glycerol) and protein inhibitor cocktail (Sigma-Aldrich). The membranes were incubated for 1 h with primary anti-Ecadherin rabbit monoclonal antibody (dilution 1/1000), anti-N-cadherin mouse monoclonal antibody (dilution 1/500), anti-vimentin rabbit monoclonal antibody (dilution 1/500), anti-phospho-AKT rabbit monoclonal antibody (dilution 1/1000), anti-phospho-ERK1/2 rabbit monoclonal antibody (dilution 1/1000), or anti-actin mouse monoclonal antibody (dilution 1/10,000) as an internal loading control, followed by 1 h with horseradish peroxidase-conjugated anti-rabbit IgG (1:5,000) or anti-mouse IgG antibody (1:20,000) (Santa Cruz Biotechnology).