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POMCDre LeprCre ROSA26lSlrSrhM3Dq and POMCDre Glp1rCre ROSA26lSlrSrhM3Dq mice and corresponding genotype controls were characterized at 13–15 weeks of age with ad libitum access to NCD. Saline (0.9%) or CNO injections (3 mg per kg body weight) were administered intraperitoneally. For measurements of food intake (as described under ‘Indirect calorimetry’), mice were injected with saline at 18:00 and 23:00, followed by a 1-d recovery period and subsequent CNO injections at 18:00 and 23:00 on the next day. For measurements of energy expenditure, respiratory exchange ratio and locomotion (as described under ‘Indirect calorimetry’), mice were injected with saline at 17:00, 22:00 and 07:00 followed by CNO at 17:00, 22:00 and 07:00 on the next day. Before perfusions at 22–26 weeks of age, mice were fasted for 2 h and injected with saline or CNO 1 h before perfusion. Serum for the corticosterone enzyme-linked immunosorbent assay (ELISA) was obtained from mice fasted for 2 h and injected with CNO 1 h before blood collection. Littermates of both sexes were used for experiments as indicated in text and figures.

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