Figure 6.

Fyn requirement for tyrosine phosphorylation of Vav induced by antagonist and agonist peptides. AD10 or Fyn-deficient AD10 T cells (1 × 10⁷) were stimulated with antagonist (T102G) pulsed, agonist (PCC 88–104) pulsed, or unpulsed DCEK.ICAM APCs (1 × 10⁷). After 1 min of stimulation, cells were lysed, immunoprecipitated with anti-Vav (A) or anti-ZAP-70 (B), and analyzed by SDS/PAGE. Proteins were transferred to membranes and immunoblotted with antiphosphotyrosine antibody (Upper). Membranes were subsequently stripped and analyzed for Vav or ZAP-70 protein (Lower). The numbers indicate the amount of phosphorylation of Vav (A) or ZAP-70 (B) relative to agonist-stimulated AD10 cells. Results are representative of three similar experiments.