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. 2020 Apr 1;37:100986. doi: 10.1016/j.molmet.2020.100986

Figure 1.

Figure 1

Measurement of real-time cAMP dynamics in pre-mature and mature murine transgenic brown adipocytes expressing Epac1-camps FRET biosensor. Single YFP and CFP channels from (A) pre-mature and (C) mature BAs expressing cytosolic FRET biosensor and their respective traces (B) and (D) followed by YFP/CFP ratio depicted as FRET traces upon stimulation with ISO (100 nM) and then by IBMX + Fsk (100 μM + 10 μM). A decrease in YFP/CFP ratio represents an increase of intracellular cAMP. (E–G) Concentration-response dependencies showing the differences in the amplitude of cAMP FRET responses induced by different β-AR ligands (E) Isoproterenol (ISO) (EC50- 92.8 nM Pre-mature BAs, 60 nM mature BAs) (F) norepinephrine (NE) (EC50- 7.7 nM Pre-mature BAs, 353.7 nM mature BAs) and (D) the β3-AR agonist CL 316,243 (CL) (EC50- 32.8 μM Pre-mature BAs, 7.8 μM mature BAs). FRET responses were normalized to basal corrected FRET ratio (R0) and the maximal response evoked by IBMX + Fsk (Rmax). n = 3-4, data are represented means ± S.E.M.