Role of Lck in CXCL12-induced chemotaxis of Lck-deficient JCaM1.6 cells.
Expression of Lck is necessary for CXCL12-induced chemotaxis of Lck-deficient JCaM1.6 cells. (A) Jurkat T cells were either left untreated or were pretreated with 1 μM HA, 1 μM PC, or 1 μM DC for 4 hours at 37°C and subjected to chemotaxis assay toward CXCL12 (25 ng/mL). Mean ± SD of 2 experiments (8 filters). (B) CXCL12-induced Lck kinase activity in JCaM1.6 cells transfected with empty vector (black bars) or with Lck wt (striped bars). A sample of Lck-transfected cells was pretreated with 100 nM DC for 4 hours. The different transfectants (4 × 107/mL) were either unstimulated or stimulated for 1 minute with CXCL12 (100 ng/mL). Mean ± SD of 3 experiments. Insert represents an immunoblot analysis of Lck in JCaM1.6 cells transfected with an empty vector or with wt Lck. (C) Chemotaxis of JCaM1.6 cells transfected with empty vector (black bars) or Lck wt (striped bars) toward CXCL12 (25 ng/mL). The Lck-transfected cells were either left untreated or pretreated with 100 nM of either PC or DC for 4 hours at 37°C. Mean ± SD of 3 experiments (18 filters). *P < .007. **P < .009. ***P < .05. P values indicate comparison with cells not treated with inhibitors.
