FIG 5.

Impact of Zta on NFAT transcription activity. The pGL3-IL2 NFAT-luciferase reporter construct was cotransfected with hisZta, a control vector, or siRNA pools into DG75 cells. Ca²⁺ signaling was activated by ionomycin-PMA immediately after transfection, with the addition of the inhibitor FK506, as indicated. After 48 h, cells were harvested for luciferase reporter and Western blot analyses. (A) The luciferase reporter promoter. The NFAT ARRE2 elements are indicated as filled boxes. (B) Luciferase assay showing the fold changes in activity after ionomycin-PMA stimulation and the impact of NFATc1 and NFATc2 siRNAs. (C) Western blot showing the impact on NFATc1 and NFATc2 expression levels. (D) Luciferase assay showing the fold changes in activity after ionomycin-PMA stimulation and FK506 inhibition and the impact of Zta expression. (E) Western blot showing Zta expression. Error bars represent the means of triplicate readings ± standard deviations. *, P < 0.05; **, P < 0.01.