Fig. 1.

NFATC1 is constitutively active in PC-3 cells. Flag-tagged NFATC1 or its mutated derivatives were transiently expressed in PC-3 prostate cancer cells. Untransfected (−) or mock-transfected cells were used as controls. a The endogenous or ectopic expression levels of NFATC1 were analysed by Western blotting with antibodies against NFATc1 or Flag, while ACTB staining was used as a loading control. b The endogenous NFAT activity of PC-3 cells was measured by luciferase assays, using transiently transfected reporters with wild-type (WT) or mutated (M) NFAT binding sites. Shown are mean luciferase activities from two independent experiments. c The effects of TPA and ionomycin on NFAT activity were measured by luciferase assays. Shown are luciferase activities of duplicate samples from one representative experiment. d Subcellular localizations of transiently expressed wild-type (WT) NFATC1, the constitutively active (mSRR) mutant and the dominant negative (DN) mutant were analysed by confocal microscopy after staining with anti-Flag antibody. Shown are average localization patterns from one experiment with three parallel samples. e The abilities of WT NFATC1 and the mSRR mutant to promote cell motility were analysed by wound healing assays from three parallel samples. Equivalent expression of these proteins was confirmed by Western blotting with anti-Flag antibody, while GAPDH staining was used as a loading control