Figure 5From: C3G forms complexes with Bcr-Abl and p38α MAPK at the focal adhesions in chronic myeloid leukemia cells: implication in the regulation of leukemic cell adhesionC3G and p38α MAPK interact with FA proteins. (A) Representative immunoprecipitation assays (IP), using the indicated antibodies, performed with protein extracts from K562 cells cultured on 10 μg/ml fibronectin for 24 h. Expression of p33, p46 and p68 phospho-paxillin (p-Pax) and paxillin isoforms, p38α, p87C3G, CrkL, Bcr-Abl, p130Cas, Abi1 and FAK was detected by immunoblotting with specific antibodies. GammaBind G Sepharose beads (B) with either buffer or whole cell lysate (lysate) were used as negative controls. (B) Confocal microscopy of K562 cells adhered for 24 h to slides covered with 10 μg/ml fibronectin and labeled with the indicated antibodies. Nuclei were labeled with DAPI. Control cells (CT) were incubated with DAPI plus the secondary antibodies. C3G-1008 (rabbit polyclonal) was used with Pax (mouse monoclonal) and p-FAK (goat polyclonal). C3G-G4 (mouse monoclonal) was used with p-Pax (rabbit polyclonal). C3G colocalizes with p-Pax and p-FAK in a punctuated pattern (white arrows). DIC: Differential interference contrast microscopy. Pax: paxillin. A488: Alexa Fluor 488. The bars represent 5 μm.Back to article page