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Calcein AM for cell viability analysis by flow cytometry Cells were seeded in 96 well plates at 3.0e5 cells/well in 100 l of RPMI-1640 with 10% FBS

AMY1R and AMY3R are the predominant subtypes expressed in the area postrema and hypothalamic nuclei relevant to energy balance regulation (Hay et al., British Journal of Pharmacology, 2015)

Cyclic AMP response element (CRE)-reporter assay A BHK cell line was stably transfected with the human calcitonin (a) receptor (hCTR a ) and a cAMP responsive element (CRE) luciferase reporter gene

Secondary antibodies were anti-Rabbit IgG 488 (Goat, 1:500, Cell Signalling #4412S), 555 (Goat, 1:500, Cell Signalling #4413S), 647 (Goat, 1:500, Cell Signalling #4414S), and 750 (Goat, 1:500, Cell Signalling #4417S) as well as anti-Rat IgG 555 (Goat, 1:500, Cell Signalling #4417S), and 647 (Goat, 1:500, Cell Signalling #4418S)