Figure 4.
NSAIDs affect TCR-dependent tyrosine phosphorylation but not CD3ζ, ZAP-70, or LAT activation. (A) Antiphosphotyrosine immunoblot of postnuclear supernatants from Jurkat cells either nonactivated (0) or activated by CD3 ligation for 5 minutes, in the presence or absence of 15 μM sc-560. After stripping, the filter was reprobed with antitubulin mAb as loading control. (B-D) Immunoblot analysis using antiphosphotyrosine mAb of CD3ζ-specific (B), ZAP-70–specific (C), or LAT-specific (D) immunoprecipitates from Jurkat cells, nonactivated or activated by CD3 ligation for 1 minute, in the presence or absence of 15 μM sc-560. The filter was stripped and reprobed with anti-CD3ζ, anti–ZAP-70, or anti-LAT antibodies as indicated. The migration of molecular mass markers is shown.