Indirect calorimetry analysis was performed at the University of Kansas Medical Center Metabolic Obesity Phenotyping Facility using a Promethion continuous indirect calorimetry system (Sable Systems International, Las Vegas, NV, USA) as previously described16. Macros available from the manufacturer were used to calculate total energy expenditure (TEE) based on the Weir equations17 and respiratory quotient (RQ) by VCO2/VO2. To calculate resting energy expenditure (REE), the Promethion-designed macro pulled out the lowest 30-min period of EE (light cycle), which was then extrapolated to a 24-h period. The difference between TEE and REE was used to determine nonresting energy expenditure (NREE). Food was weighed before and after each experiment to determine food consumption. The mass of food consumed was used to determine energy intake (EI) based on the manufacturer-provided values for metabolizable energy from each diet type, and comparisons were made using body mass as a covariate for EI. Energy balance (EB) was calculated by subtracting daily TEE from EI. These calculations were performed as previously described18. Food hoppers were equipped with a sensor to determine food hopper interaction, and manufacturer-designed macros used these data to determine hourly feeding bouts. Activity was determined using the XYZ Beambreak Activity Monitor (Sable Systems) and is expressed as the sum of X and Y beam breaks per time period. Mice were given 2 days to acclimatize to new cages before data were recorded.

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