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. 2000 Feb 21;191(4):651–660. doi: 10.1084/jem.191.4.651

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Effects of anti-CD3/CD40L costimulation on apoptosis and expression of cell survival proteins. (A) Viability kinetics in anti-CD3/CD40L– and anti-CD3/CD28–stimulated cultures. Trypan blue uptake was assessed in cell suspensions. (B) Apoptosis in cultured cells was measured using the TUNEL method to measure dUTP incorporation into single-strand DNA breaks. CD4⁺ T cells irradiated with 1,000-Gy γ rays 24 h previously served as a positive control for the assay. Values represent the mean ± SEM from five individual experiments. (C) Expression of the cell survival gene Bcl-xL and Bcl-2. Protein lysates from 10⁷ cells stimulated for 72 h were subjected to sequential immunoprecipitation with a Bcl-xL–specific mAb and immunoblotted with a Bcl-xL rabbit polyclonal antibody as described.

Effects of anti-CD3/CD40L costimulation on apoptosis and expression of cell survival proteins. (A) Viability kinetics in anti-CD3/CD40L– and anti-CD3/CD28–stimulated cultures. Trypan blue uptake was assessed in cell suspensions. (B) Apoptosis in cultured cells was measured using the TUNEL method to measure dUTP incorporation into single-strand DNA breaks. CD4⁺ T cells irradiated with 1,000-Gy γ rays 24 h previously served as a positive control for the assay. Values represent the mean ± SEM from five individual experiments. (C) Expression of the cell survival gene Bcl-xL and Bcl-2. Protein lysates from 10⁷ cells stimulated for 72 h were subjected to sequential immunoprecipitation with a Bcl-xL–specific mAb and immunoblotted with a Bcl-xL rabbit polyclonal antibody as described.