Fig. 8. Dab2 is transiently phosphorylated and translocates into the cytoskeleton/membrane fraction in response to adhesion. (A) RAW macrophages overexpressing Dab2 were trypsinized and plated on laminin-coated culture dishes or were kept in suspension. At the time points indicated, total cell extracts of adherent cells and cells kept in suspension were analyzed by western blotting. RAW cells overexpressing Dab2 (B) or ICSBP^–/– BMM (C) were treated as described in (A), and plated on culture dishes coated with fibronectin (FN), poly-d-lysine (pDL) or laminin (LA). After 30 min, total cell extracts of adherent cells and cells kept in suspension were prepared, and subjected to SDS–PAGE and western blotting with the anti-Dab2 antibody. (D) RAW cells overexpressing Dab2 were trypsinized and plated on culture dishes coated with fibronectin, or kept in suspension. After 60 min, cytosolic and particulate fractions were prepared as described in Materials and methods, and were analyzed by western blotting with anti-Dab2 antibody.