Figure 4

P-FAK and p-PI-3K expression in cultured CTC from breast cancer patients. (A) Proteins obtained from breast cancer patients and normal female blood donors cultured cells were immunoprecipitated (IP) with a phosphotyrosine (p-Tyr) antibody. FAK was identified by western blot analysis using an anti-FAK polyclonal antibody. Right panels: Representative micrographs of confocal laser scanning microscopic sections of cultured PBMC stained with pancytokeratins polyclonal antibody (red a), p-FAK monoclonal antibody (green b), and overlay section (c). Magnification, ×600. (B) As described in for section A. PI-3 kinase was identified by western blot analysis using an anti-PI-3 kinase polyclonal antibody. Right panel: Representative micrographs of confocal laser scanning microscopic sections of cultured PBMC stained with pancytokeratins monoclonal antibody (green a), p-PI-3K polyclonal antibody (red b), and overlay section (c). Magnification, ×600. (C) p-FAK and CK expression in cultured CTC after immunomagnetic separation with EpCAM antibody and western blot analysis with p-FAK and CK antibodies respectively. Note that CD45 is not expressed.