Figure 3
Cytoskeletal and focal adhesion changes. (A) Phosphorylation status of the cytoskeletal regulatory proteins p130CAS, Crk, and paxillin following drug treatments. Immunoblot analysis indicates down-modulation of cytoskeletal regulatory proteins in terms of protein levels (only p130CAS) and tyrosine phosphorylation following dasatinib treatment. (B) Confocal microscope images of TPC-1 cells before and after drug treatments. Cells were stained for phospho-paxillin (red), actin (green phalloidin) and DRAQ5 to highlight the focal adhesions. In control and RPI-1-treated cells, phospho-paxillin accumulated in well-defined zones around the focal adhesions, while after dasatinib treatment, phospho-paxillin disappeared. Confocal images (512 × 512 pixels) were obtained using a 60× oil immersion lens and analyzed using ImagePro 6.3 software. Scale bars, 10 μm.