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Figure 6 | Journal of Molecular Signaling

Figure 6

From: AKAR2-AKAP12 fusion protein "biosenses" dynamic phosphorylation and localization of a GPCR-based scaffold

Figure 6

FRET-based " biosensing" by AKAR2(T/P) mutant-AKAP12 in response to beta-adrenergic agonist. Stably transfected A431 clones expressing AKAR2(T/P) mutant of AKAP12 (lacking the phosphorylation site for PKA in the substrate domain of the AKAR2 moiety) were serum starved over night and then untreated (-ISO) or treated with beta-adrenergic agonist (isoproterenol, 10 μM for 10 min (+ ISO 10 min). Upper panel, images and fluorescent densities recorded for CFP and YFP are displayed. The measurements were made in the perinuclear, cytoplasmic area (open red circle), an area in which robust FRET signals from activated AKAR2-AKAP12 are observed. Lower panel, the fluorescence density measurements were analyzed from a large array of images collected for this purpose. The fluorescence density of CFP in the same area of the cells at basal state (-ISO) is highlighted by a red dashed line.

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