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

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Combined triggering of CD3 and CD28 delays downregulation of ERK2 activity by CTLA-4. Preactivated T cells were coated with different combinations of antibodies as in Fig. 1 and lysed 1, 5, or 10 min after addition of warm cross-linking antibody. ERK2 was immunoprecipitated and in vitro kinase reactions were performed (top). The same lysates were tested for ERK2 protein abundance (middle). ERK2 activities were quantified using a phosphorimager and represented as relative values compared to ERK2 activities from unstimulated cells (unstimulated = 1; bottom).

Combined triggering of CD3 and CD28 delays downregulation of ERK2 activity by CTLA-4. Preactivated T cells were coated with different combinations of antibodies as in Fig. 1 and lysed 1, 5, or 10 min after addition of warm cross-linking antibody. ERK2 was immunoprecipitated and in vitro kinase reactions were performed (top). The same lysates were tested for ERK2 protein abundance (middle). ERK2 activities were quantified using a phosphorimager and represented as relative values compared to ERK2 activities from unstimulated cells (unstimulated = 1; bottom).