Association of Pyk2 and Syk with β2 integrin in fMLP-stimulated granulocytic cells.
Granulocytic cells were incubated for the indicated times with 1 μmol/L fMLP on Fg-coated dishes (attached) or in suspension in polypropylene tubes (suspension). Lysates from the equivalent number of cells (5 × 106 cells) were immunoprecipitated with anti-β2 integrin monoclonal antibody, normal mouse IgG, protein A alone (A-B), anti-Pyk2 polyclonal antibody (A), or anti-Syk (SC-929) polyclonal antibody (B). Immunoblotting analysis was performed with anti-Pyk2 (A) or anti-Syk (B) polyclonal antibody. Each blot was stripped and reprobed with anti-β2 integrin polyclonal antibody (A-B, lower panels). The positions of molecular markers are shown on the left in kilodaltons. Arrows indicate the position of a Pyk2 (A, upper panel), Syk (B, upper panel), or β2 integrin (A-B, lower panels) protein band.
