Effects of wortmannin, GF109203X, or PD98059 on the SDF-1–induced tyrosine phosphorylation of various adhesion proteins in CTS cells. Serum-starved CTS cells were incubated without or with 100 nmol/L wortmannin, 3 μmol/L GF109203X, or 20 μmol/L PD98059 (PD) for 45 minutes, then stimulated with SDF-1α (20 nmol/L) or left untreated for 1 minute at 37°C. Total cell lysates were immunoprecipitated with antibodies for FAK (A), RAFTK (B), p130 Cas (C), paxillin (D), Crk (E), or Crk-L (F), respectively. The immunoprecipitates were subjected to immunoblot analysis with anti-PY antibody (4G10) (upper panels), followed by reprobing with the same antibody used for each of the specific immunoprecipitations (lower panels). Lanes 1 to 8 represent the control, untreated cells with SDF-1α stimulation (SDF-1α), wortmannin-treated cells without (Wort.) or with SDF-1α (Wort.+SDF-1α) stimulation, GF109203X-treated cells without (GF) or with SDF-1α (GF+SDF-1α) stimulation, PD98059-treated cells without (PD) or with SDF-1α (PD+SDF-1α) stimulation, respectively. The increase in tyrosine phosphorylation of each adhesion protein is indicated (by fold increase) based on the densitometry values.