Fig. 9. Anandamide-induced Ca²⁺ signal subsequently activates NFAT, Erk1 and Erk2. (A,B) The effect of 10 µM anandamide on the (trans)location of GFP-tagged NFAT was visualized in endothelial cells in the presence (A) or absence (B) of 2 mM extracellular Ca²⁺. (C,D) Time dependency of the effect of 10 µM anandamide on Erk1 and Erk2 phosphorylation in endothelial cells in 2 mM Ca²⁺-containing buffer in the absence (C) or presence (D) of 70 µM Mn²⁺. Images of endothelial cells that were transiently transfected with NFAT-GFP were taken 48 hours after transfection using a conventional fluorescence microscope. Data presented are representative for at least eight cells out of three experiments on alternate days. Scale bars: 10 µm. For assessment of Erk1 and Erk2 activation, standard western blotting was applied.