Figure 2. CD39 expression in CD8⁺ T cells is JNK and NFκB dependent.

(a,b) Healthy blood CD8⁺ T cells were treated with anti-CD3 (10 μg ml⁻¹, precoated) and anti-CD28 (5 μg ml⁻¹, soluble) antibodies, or 10 ng ml⁻¹ of the cytokines: either TNF or IL-12. CD39 expression was then determined by flow cytometry at 24 h (a) (n=4) or by quantitative PCR at 2 h (b) (n=4). (c) Healthy blood CD8⁺ T cells were stimulated with anti-CD3/CD28 antibodies in the presence or absence of VAS2870 (10 μM), NAC (10 mM), JNK inhibitor II (10 μM), PS-1145 (10 μM) for 72 h. CD39 expression was then analysed by fluorescence-activated cell sorting (n=3). (d,e) Chromatin immunoprecipitation analyses indicating enrichment of NFκB p65 (d) or JNK/c-Jun (e) at the CD39 promoter region in CD8⁺ T cells fresh isolated (fresh) or activated with anti-CD3/28 antibodies for 24 h (activated). Data are shown as mean±s.e.m., **P<0.001 (one-way analysis of variance), for the comparison with the other groups.