Fig. 2. Interaction between EphA1 and ILK in mammalian cells. (A) Cells expressing GFP, WT (wild-type)-EphA1–GFP or KD (kinase dead)-EphA1–GFP were transfected with ILK-myc. 24 hours after transfection, cells were serum-starved for 12 hours and stimulated with ephrin-A1–Fc (1 µg/ml) for 10 minutes. Cell lysates were immunoprecipitated (IP) with anti-myc antibody followed by immunoblot (IB) analysis with the indicated antibodies. (B) HEK293 cells expressing WT-EphA1–GFP or SAM-EphA1–GFP were transfected with ILK-myc and treated as in A. Whole cell lysate (WCL; lane 1) and immunoprecipitates by control IgG (lane 2), anti()-GFP (lanes 3-6), anti-myc (lane 7) and no antibody (Ab) (lane 8) were subjected to anti-EphA1 and anti-ILK immunoblotting. (C) EBC-1 cells were serum-starved overnight and stimulated with ephrin-A1–Fc (1 µg/ml) for 10 minutes. Cell lysates were immunoprecipitated with anti-ILK antibody followed by immunoblot analysis with anti-EphA1 and anti-ILK antibody.