Sensitivity of NFAT5 transcriptional activity and expression to pharmacological inhibitors. A) Transgenic 9xNFAT-Luc splenocytes preactivated during 24 hours with concanavalin plus IL2 were treated with hypertonicity (400 mOsm/kg, 24 hours) in the absence or presence of FK506 (100 nM), SB203580 or SB202190 (both at 10 μM), PD98059 (10 μM), H89 (2 μM), wortmannin (0.5 μM), or LY294002 (LY). Luciferase activity in the upper panel (mean ± S.D of three independent experiments) is shown as percentage of the activity in the absence of inhibitors (100%). Western blotting in the lower panelshows NFAT5 and pyruvate kinase expression. B) Transgenic MEF were stimulated for 8 or 24 hours with hypertonicity (460 mOsm/kg) in the absence or presence of SB203580 (10 μM) or LY294002 (LY). Luciferase activity is shown is the upper panel (mean ± S.E.M of three independent experiments). NFAT5 and pyruvate kinase expression are shown in the lower panel.