Figure 16 Isoproterenol stimulation of Jurkat cells transfected with NF-AT-β-lactamase vector. The upper panel shows fluorescence micrographs of individual wells seeded with Jurkat cells after 4 h incubation in the dual-FRET fluorophore CCF2. The pair of panels of green cells at the left show cells that were not treated with isoproterenol. The green fluorescence originates by direct excitation of the coumarin and sensitization of fluorescein fluorescence by FRET in the intact CCF2 molecule. The leftmost panel was obtained by seeding 250 cells in the well, whereas the adjacent panel to the right shows 1000 cells. The plot at bottom shows the quantitative relation between blue (coumarin) and green (fluorescein) fluorescence intensities as a function of cell density. In unstimulated cells, green fluorescence predominates. The pair of panels at the right shows Jurkat cells after stimulation with isoproterenol. The expressed β-lactamase has cleaved the dual-FRET fluorophore, relieving the quenching of the coumarin fluorescence. See color plate.

doi:10.1201/9780203910696-89

Chapter

Figure 16 Isoproterenol stimulation of Jurkat cells transfected with NF-AT-β-lactamase vector. The upper panel shows fluorescence micrographs of individual wells seeded with Jurkat cells after 4 h incubation in the dual-FRET fluorophore CCF2. The pair of panels of green cells at the left show cells that were not treated with isoproterenol. The green fluorescence originates by direct excitation of the coumarin and sensitization of fluorescein fluorescence by FRET in the intact CCF2 molecule. The leftmost panel was obtained by seeding 250 cells in the well, whereas the adjacent panel to the right shows 1000 cells. The plot at bottom shows the quantitative relation between blue (coumarin) and green (fluorescein) fluorescence intensities as a function of cell density. In unstimulated cells, green fluorescence predominates. The pair of panels at the right shows Jurkat cells after stimulation with isoproterenol. The expressed β-lactamase has cleaved the dual-FRET fluorophore, relieving the quenching of the coumarin fluorescence. See color plate.

Chapter

Figure 16 Isoproterenol stimulation of Jurkat cells transfected with NF-AT-β-lactamase vector. The upper panel shows fluorescence micrographs of individual wells seeded with Jurkat cells after 4 h incubation in the dual-FRET fluorophore CCF2. The pair of panels of green cells at the left show cells that were not treated with isoproterenol. The green fluorescence originates by direct excitation of the coumarin and sensitization of fluorescein fluorescence by FRET in the intact CCF2 molecule. The leftmost panel was obtained by seeding 250 cells in the well, whereas the adjacent panel to the right shows 1000 cells. The plot at bottom shows the quantitative relation between blue (coumarin) and green (fluorescein) fluorescence intensities as a function of cell density. In unstimulated cells, green fluorescence predominates. The pair of panels at the right shows Jurkat cells after stimulation with isoproterenol. The expressed β-lactamase has cleaved the dual-FRET fluorophore, relieving the quenching of the coumarin fluorescence. See color plate.

ABSTRACT

Figure 16 Isoproterenol stimulation of Jurkat cells transfected with NF-AT-β-lactamase vector. The upper panel shows fluorescence micrographs of individual wells seeded with Jurkat cells after 4 h incubation in the dualFRET fluorophore CCF2. The pair of panels of green cells at the left show cells that were not treated with isoproterenol. The green fluorescence originates by direct excitation of the coumarin and sensitization of fluorescein fluorescence by FRET in the intact CCF2 molecule. The leftmost panel was obtained by seeding 250 cells in the well, whereas the adjacent panel to the right shows 1000 cells. The plot at bottom shows the quantitative relation between blue (coumarin) and green (fluorescein) fluorescence intensities as a function of cell density. In unstimulated cells, green fluorescence predominates. The pair of panels at the right shows Jurkat cells after stimulation with isoproterenol. The expressed β-lactamase has cleaved the dual-FRET fluorophore, relieving the quenching of the coumarin fluorescence. See color plate.