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. 2018 May 8;11:148. doi: 10.3389/fnmol.2018.00148

FIGURE 3.

FIGURE 3

Inhibition of AC reduces the L-lactate- and 3Cl-5OH-BA-induced increase in [cAMP]i and [lactate]i (A,B, panels i) Mean time-courses of FRET ratio signal changes normalized to the maximum signal change for (A,i) Epac1-camps upon addition of 20 mM L-lactate and (B,i) Laconic upon addition of 0.5 mM 3Cl-5OH-BA (black lines) in the absence (white circles) and presence (black circles) of 100 μM DDA, an inhibitor of AC. Each data point represents mean ± s.e.m. (A,B, panels ii) Mean relative changes in FRET ratio (Rel. ΔFRET ratio) upon L-lactate (A) and (B) 3Cl-5OH-BA stimulation in the absence and presence of DDA. Relative ΔFRET values (%) were calculated by dividing individual ΔFRET values with the average ΔFRET value upon L-lactate or 3Cl-5OH-BA stimulation. Note that the inhibition of AC by DDA causes ∼50 % reduction in L-lactate-induced increase in [cAMP]i in astrocytes and a ∼30–60% reduction in 3Cl-5OH-BA-induced increase in [lactate]i in astrocytes, 3T3-L1 and BT474 cells. In BT474 cells the application of 3Cl-5OH-BA initiated a transient reduction in [lactate]i that was diminished in the presence of DDA. Numbers adjacent to black bars represent number of cells. Data are in the format of the mean ± s.e.m (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001). Data for every set of experiment was acquired from at least two different animals.