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cAMP accumulation assay FlpInCHO-hGLP-1R and FlpInCHO-pGLP-1R cells were diluted to the density of 6 10 5 cells/ml and INS-1(832/13) cells to 2 10 6 cells/ml in stimulation buffer [phenol-free DMEM containing 5 mM HEPES, 0.5 mM 3-isobutyl-1-methylxanthine (IBMX) and 0.1% (w/v) BSA], and cAMP detection was carried out as described previously 48

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Samples were analyzed on at least three representative 1.5-mm cores taken from each donor block following the selection of regions of interest by a pathologist

To further support a role for GLP-1R signaling in regulating food reward, recent fMRI studies in obese and obese T2DM patients have demonstrated that intravenous infusion of exenatide decreases the activation of brain regions involved in mediating food reward (van Bloemendaal et al., 2014)