Coronary stent implantation is often followed by in-stent restenosis. This re-narrowing of the vessel results from the formation of neointima, which is mainly composed of proliferative smooth muscle cells. In the stenting procedure, the vessel wall is subjected to mechanical strain, which might promote cell proliferation. Stretching the cell membrane is considered to trigger Ca2+ influx through stretch-activated channels, thereby activating Ca2+ dependent transcription factors that promote cell proliferation, such as the nuclear factor of activated T-cells (NFAT). We hypothesized that transient receptor potential canonical (TRPC) channels and/or store-operated Orai channels constitute Ca2+ entry pathways sensitive to mechanical stress and linked to NFAT activation.NFAT activation was characterized in vascular cells in response to receptor-stimulation, store-depletion, and stretch, as well as NFAT inhibition by the pyrazole compound Pyr3. Store-operated Ca2+ entry, induced by thapsigargin, effectively activated NFAT, which indicates that store-operated Ca2+ channels are involved in NFAT activation. Pyr3 potently inhibited NFAT activation in vascular smooth muscle, as well as in endothelial cells. Pyr3 was originally proposed to be a selective TRPC3 inhibitor. However, recent findings suggest that Pyr3 generally inhibits store-operated Ca2+ entry.Cultured vascular smooth muscle cells were stretched on flexible membranes to explore whether NFAT activation could be triggered by single static stretch. Stretching experiments with porcine aortic smooth muscle cells implied stretch-mediated NFAT activation. Human aortic smooth muscle cells did not display significant NFAT activation in response to stretch. The results obtained so far argue against a sensitivity of human vascular smooth muscle NFAT signaling to a single, tonic increase in mechanical stress. Further experiments are needed to elucidate the mechanisms of mechanosensation in human vascular smooth muscle.