Fig. 5. Recruitment of Shc and Grb2. (A) CHO_Vß₃ cells and (B) osteoclast cultures were treated as described in Fig. 2, immunoprecipitated with the indicated antibodies and western blotted to detect PKC, Shc (diluted 1:200), Grb2 (diluted 1:1000) and ß₃ integrin (left and middle panels), or to detect total tyrosine-phosphorylated proteins using the PY99 (diluted 1:1000) antibody (right panels). Similar results were obtained in three independent experiments. IP, immunoprecipitation; TCL, total cell lysate; WB, western blot analysis; Susp, cells in suspension. LM609, adhesion to LM609 for 30 minutes.