rise in human subcutaneous fibroblasts depends on B
receptor activation. Calcium oscillations in human fibroblasts of the subcutaneous tissue stimulated with bradykinin (BK, 30 μM) were tested after pretreatment of the cells with selective B1 and B2 receptor antagonists, respectively R715 (1 μM, A) and HOE-140 (1 μM, B; 10 μM, C). Cells were pre-incubated with the cell-permeant fluorescent calcium indicator, Fluo-4 NW (2.5 μM, see Methods). Changes in fluorescence were detected using a microplate reader. Intracellular Ca2+ transients were calibrated to the maximal calcium load produced by ionomycin (5 μM, 100% response). Black arrows indicate the time of drugs application. No changes in baseline fluorescence were observed after application of the antagonists. Each point represents pooled data from an n number of experiments. The vertical bars represent S.E.M.. *p < 0.05 represent significant differences from BK (30 μM) alone.