Firefly luciferase constructs downstream of promoters for NF-kB or NF-AT were co-transfected with renilla luciferase pRL-TK (Int -) plasmid (Promega). Luciferase activities were analyzed using the Dual-Luciferase Kit (Promega), with firefly fluorescence units normalized to renilla luciferase fluorescence units (BMG microplate reader). were purified from blood on Ficoll-isopaque gradients. PBL were nucleofected with the Nucleofactor system and T cell solution (Amaxa, program U14), and left for three days in culture medium prior treatment.