Fig. 8. SHIP2 associates with Shc via the NPXY motif. (A) Anti-SHIP2 or control (Pre) IPs from HeLa cells that were detached (D) or replated for 60 minutes on collagen I (6 µg/cm²; C.I) were blotted with anti-Shc (monoclonal) or anti-SHIP2 antibodies. (B) Anti-SHIP2 or control (Pre) IPs, from HeLa cells that were re-plated for 60 minutes on collagen I (6 µg/cm²; C.I), fibronectin (5 µg/cm²; F), or poly-L-lysine (0.01% solution, 0.5 ml/25 cm²; P-Ly) were blotted with anti-Shc (monoclonal) or anti-SHIP2 antibodies as indicated. Cells were plated in the presence of vehicle control DMSO (-) or 1 µM Src inhibitor compound PD180970 (+). Whole cell lysate was used as a control (WCL). (C) HeLa cells were transiently transfected with wild-type or YY-FF (986-987) mutant SHIP2 expression constructs. 48 hours post-transfection, serum-starved cells were re-plated on collagen I (6 µg/cm²) for 60 minutes. Anti-FLAG (rabbit polyclonal) IPs from these samples were blotted with anti-Shc (monoclonal) or monoclonal anti-FLAG (M2) antibodies. Whole cell lysate was used as a control (WCL). (D) Anti-Shc (monoclonal) or control mouse IgG (MIg) IPs from HeLa cells re-plated for 60 minutes on collagen I (6 µg/cm²; C.I), fibronectin (5 µg/cm²; F), or poly-L-lysine (0.01% solution, 0.5 ml/25 cm²; P-Ly) were blotted with anti-SHIP2 or anti-Shc (polyclonal) antibodies as indicated. Cells were plated in the presence of either vehicle control DMSO (-) or 1 µM Src inhibitor compound PD180970 (+). Whole cell lysate was used as a control (WCL). Small arrows point to the three forms of Shc proteins. The large arrowhead points to SHIP2 protein.

Return to article