FIG. 3.

Binding of GSTnef with TCR_ζ in vitro. GST, recombinant GST239nef, and GSTSF2nef proteins were expressed in E. coli, affinity purified, and immobilized by using glutathione-Sepharose beads. The immobilized GST proteins were incubated with extracts of Cos 18 or Jurkat cells previously precleared with immobilized GST and glutathione-Sepharose beads. After extensive washing, proteins coprecipitated with the complexes were analyzed by SDS-12% PAGE, electroblotted onto a nylon membrane, and reacted with anti-TCR_ζ monoclonal antibody.