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

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CTLA-4 reduces JNK activity induced by CD3 and CD28 triggering. Preactivated T cells were coated with different combinations of antibodies and lysed 10 min after addition of warm cross-linking antibody. Lysates were tested for JNK activity by precipitation with GST–c-jun precoupled to glutathione beads followed by in vitro kinase reactions as described in Materials and Methods. Phosphorylated GST–c-jun is indicated by the arrow (top). The same lysates were tested for JNK protein abundance on immunoblot (middle). JNK activities were quantified using a phosphorimager and represented as relative values compared to JNK activity from unstimulated cells (unstimulated = 1; bottom). Data for the 10-min time point are shown because no JNK activity could be measured at earlier times.

CTLA-4 reduces JNK activity induced by CD3 and CD28 triggering. Preactivated T cells were coated with different combinations of antibodies and lysed 10 min after addition of warm cross-linking antibody. Lysates were tested for JNK activity by precipitation with GST–c-jun precoupled to glutathione beads followed by in vitro kinase reactions as described in Materials and Methods. Phosphorylated GST–c-jun is indicated by the arrow (top). The same lysates were tested for JNK protein abundance on immunoblot (middle). JNK activities were quantified using a phosphorimager and represented as relative values compared to JNK activity from unstimulated cells (unstimulated = 1; bottom). Data for the 10-min time point are shown because no JNK activity could be measured at earlier times.