Activation of the 9xNFAT-luc reporter by NFATc or NFAT5. A) Jurkat T cells transfected with the 9xNFAT-Luc reporter were stimulated with PMA plus ionomycin (P+I) or cultured in hypertonic medium (500 mOsm/kg) without or with FK506 during 24 hours. Luciferase activity is represented as relative light units per second (RLU) after normalization with Renilla and endogenous lactate dehydrogenase. Mean ± S.D of four independent experiments is shown. B) 9xNFAT-Luc and vectors encoding GFP, the NFAT5-inhibitory dimerization domain (DD5-GFP) or the NFATc-inhibitory peptide VIVIT (VIVIT-GFP) were transfected in Jurkat T cells. Cells were treated during 24 hours with PMA plus ionomycin or hypertonicity (500 mOsm/kg) in the absence or presence of FK506. Mean ± S.D of three independent experiments is shown. N.S.: non statistically significant. C) Jurkat T cells transfected with the 9xNFAT-Luc reporter or the ORE-luc reporter were exposed to increasingly hypertonic media in the presence of FK506, or stimulated with PMA plus ionomycin without FK506. Mean ± S.D of three independent experiments is shown.