In vitro functionality of NFAT-inducible lentiviral vector. (A) 293T and 3T3 cell lines were transfected with the NFAT vector before stimulation with PMA (100 ng/mL)/ionomycin (1500 ng/mL) for 6 hours in the presence or absence of CsA (300 ng/mL). NFAT-CBRLuc activity normalized to extGLuc activity was measured using a dual luciferase assay and expressed as a fold induction above unstimulated cells. Data shown represent averages ± SEM. 293T, n = 8; 3T3, n = 6. *P < .05, stimulated cells vs control. (B) EL4 and Jurkat cells were transduced with the NFAT vector. Three days after transduction cells were stimulated overnight with PMA (100 ng/mL)/ionomycin (1500 ng/mL) and αCD3/CD28 for 16 hours in the presence or absence of CsA (300 ng/mL). Luciferase activity was detected by using the Dual-Glo Luciferase Assay System (Promega) in a Clarity Luminescence Microplate Reader (BioTek) or using the IVIS bioluminescence imaging system (Xenogen). Data shown represent averages ± SEM. EL4, n = 6; Jurkat, n = 5. *P < .05, stimulated cells vs control.