Nec-1 blocks sustained phosphorylation of LAT during T cell activation.

Abstract

<p>(A) Whole cell extracts (WCE) were examined from T cells stimulated with anti-CD3 antibody in the presence or absence of 50 µM Nec-1. Western blot was performed with phospho-tyrosine specific antibody. Short and long exposures were shown to highlight the different phosphorylated species. (B) Nec-1 impairs TCR-induced LAT phosphorylation. Total lysates from activated T cells were subjected to immunoprecipitation with phospho-tyrosine antibody. The resulting immune complex was evaluated for presence of LAT and Lck by Western blot. (C) Nec-1 does not affect Erk phosphorylation. Purified T cells were stimulated as in (A). Western blot was performed with phospo-Erk or total Erk specific antibodies.</p