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. 1997 Nov 17;186(10):1645–1653. doi: 10.1084/jem.186.10.1645

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Anti-CD3–induced phosphorylation of TCR-ζ and ZAP70 is not affected by CTLA-4. (A) Preactivated T cells were coated with different combinations of antibodies and lysed 1, 5, or 10 min after addition of warm cross-linking antibody. TCR-ζ was immunoprecipitated and immunoblotted with antiphosphotyrosine antibody 4G10. Indicated with arrows are the p21 and p23 isoforms of tyrosine phosphorylated TCR-ζ. (B) Lysates prepared as in A from cells stimulated for 1 min were immunoprecipitated with anti-ZAP70 antibody and immunoblotted with antiphosphotyrosine antibody 4G10. Indicated with an arrow is phosphorylated ZAP70. Similar results were also obtained on lysates from 5 and 10 min–stimulated cells (not shown).

Anti-CD3–induced phosphorylation of TCR-ζ and ZAP70 is not affected by CTLA-4. (A) Preactivated T cells were coated with different combinations of antibodies and lysed 1, 5, or 10 min after addition of warm cross-linking antibody. TCR-ζ was immunoprecipitated and immunoblotted with antiphosphotyrosine antibody 4G10. Indicated with arrows are the p21 and p23 isoforms of tyrosine phosphorylated TCR-ζ. (B) Lysates prepared as in A from cells stimulated for 1 min were immunoprecipitated with anti-ZAP70 antibody and immunoblotted with antiphosphotyrosine antibody 4G10. Indicated with an arrow is phosphorylated ZAP70. Similar results were also obtained on lysates from 5 and 10 min–stimulated cells (not shown).