Figure 1

Cytokeratin and p-FAK expression in CTC from breast cancer patients. (A) Representative micrograph of cytospin, double stained with CD45 monoclonal antibody (green) for hematopoietic cells and with pancytokeratins polyclonal antibody (red) for circulating tumor cells. Magnification, ×400. Right panel: Quantification of CK-positive samples by immunofluorescence microscopy and RT-PCR analysis. (B) Representative micrograph of PBMC cytospin stained with pancytokeratins polyclonal antibody (red), p-FAK monoclonal antibody (green), and DAPI (blue). Magnification, ×400. (C) Confocal laser scanning microscopy analysis of PBMC cytospins stained with p-FAK monoclonal antibody (green) and pancytokeratins polyclonal antibody (red). Magnification, ×600. Right panel: Quantification of p-FAK and CK co-expression by immunofluorescence microscopy in CTC. (D) Representative micrograph of PBMC cytospins double stained with FAK polyclonal antibody (red) for total protein and with p-FAK monoclonal antibody (green) for activated kinase. Note that FAK is expressed in all the cells, while pFAK is expressed only in a distinct cell. Magnification, ×200. (E) p-FAK expression in CTC after immunomagnetic separation of PBMC with Ep-CAM antibody coated beads and indirect immunofluorescence with pancytokeratins polyclonal antibody (red), p-FAK monoclonal antibody (green), and DAPI (blue). Magnification, ×600.