FIGURE 5.

Focal adhesion kinase is involved in the regulation of syndecan-2-mediated melanoma cell migration. A, B16 cells transfected with either the empty vector (vec) or sdc2 cDNA were lysed with radioimmune precipitation buffer and analyzed by Western blotting with anti-phosphotyrosine (α-PY) antibody (left). The same membrane was stripped and reprobed with antibodies against FAK (α-FAK), paxillin (α-paxillin), or Erk (α-Erk). B, B16 cells were lysed, and site-specific FAK phosphorylation was analyzed with anti-FAK(Tyr(P)³⁹⁷) and anti-FAK(Tyr(P)⁸⁶¹) antibodies, followed by stripping and reprobing with anti-FAK antibody. C, B16 cell extracts were immunoprecipitated with an antibody against paxillin, and the phosphorylation state of proteins was determined by immunoblotting anti-phosphotyrosine (PY) antibody. The levels of protein in each immunoprecipitate were determined by immunoblotting with anti-paxillin antibody.