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. Author manuscript; available in PMC: 2012 Dec 15.
Published in final edited form as: J Immunol. 2011 Nov 9;187(12):6197–6207. doi: 10.4049/jimmunol.1101348

Figure 2. Deletion of CARMA1 after T cell activation reduces T cell reactivation in vitro.

Figure 2

A) CD4+ T cells isolated from mice were stimulated with anti-CD3 and anti-CD28 coated antibodies, rested and reactivated. B and C) Transcript expression levels of CARMA1 (B) and OX40 (C) on days 8 and 9 of the experimental protocol, as analyzed by real-time quantitative PCR. Data from 3 mice per genotype are shown as mean ± SEM. *p<0.05 by t-test. D) Representative flow cytometry plots demonstrating CD69 and CD44 expressing CD4+ T cells without (−) or with (+) anti-CD3 and anti-CD28 restimulation, assessed on day 9. All plots depict events in live lymphocyte gate. E) Percentage of activated (CD4+CD69+) and memory-phenotype (CD4+CD44+) T cells without (−) or with (+) anti-CD3 and anti-CD28 restimulation, assessed on day 9. Data reported as mean ± SEM from 9–10 mice in 4 separate experiments. *p<0.05 by t-test. F) Nuclear translocation of NF-κB p65 subunit in CD4+ T cells without (−) or with (+) anti-CD3 and anti-CD28 restimulation. Each lane was loaded with 5 µg of nuclear protein extracted from cells 2 hours following restimulation. Following immunoblotting for CARMA1, membranes were stripped and reprobed for Lamin-B to ensure equal nuclear protein loading. Blot representative of 2 independent experiments with 2 different samples per genotype.