Figure 5

Imaging PKA-catalyzed phosphorylation via AKAP5 versus AKAP12: response to stimulation of the cells by β-adrenergic agonist. The YFP/CFP emission ratio images of HEK 293 cells was performed in cells expressing either AKAR2, AKAR2-AKAP5, or AKAR2-AKAP12. A time = 0, isoproterenol (ISO) was applied to the cultures. The first images displayed for each array are YFP-only images. Representative pseudocolor images display the FRET signal (PKC-catalyzed phosphorylation of the AKAR2 and two AKAP biosensors in response to ISO (10 μM). The sampling was performed at the time points indicated, recorded at every 5 s for a period of 2 min using METAFLUOR software. Experiments were repeated three times with equivalent results.