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Glucose tolerance tests were performed at 13 weeks of age with 16-h fasted mice. Body weights of mice and their basal blood glucose using a glucometer and glucose strips (Contour Next, Bayer HealthCare) were determined before the start of the experiment. Mice were injected with 20% glucose, and blood glucose was measured at 15, 30, 60 and 120 min after injection.

Lean and fat mass were determined via nuclear magnetic resonance (NMR Analyzer Minispec mq 7.5, Bruker Optik) in live mice. Alternatively, body composition was analyzed by computed tomography (CT) in isoflurane-anesthetized mice (Dräger and Piramal Healthcare). For data acquisition on an IVIS Spectrum CT scanner (Caliper LifeScience), we used IVIS LivingImage Software V.4.3.1. Quantification of lean and fat mass contents were determined with a modification of the previously described VINCI software package 4.61.0.

Metabolic phenotyping and food intake were measured by an automated PhenoMaster open-circuit indirect, calorimetry system (TSE Systems). Mice were allowed to acclimatize to the experimental setup for 4 d before the start of each experiment. Food and water were available ad libitum. Data acquisition was carried out by TSE Phenomaster versions 6.2.5 and above.

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