Whole body composition was monitored using Echo MRI-100 whole-body composition analyzer (Echo Medical Systems, Houston, TX, USA) to assess fat and lean mass. All metabolic parameters data were obtained using an Oxymax (Columbus Instruments, Columbus, OH, USA) open-circuit indirect calorimetry system, as we have previously described [11 (link),71 (link)]. Mice were individually housed in feeding chambers and given free access to powdered diets for 4 days for initial acclimatization, then transferred to metabolic chambers for indirect calorimetry testing for 5 days. Data presented are averages of the day 2 to 4 of indirect calorimetry recordings. On the last day, the mice were fasted starting from 6 a.m. (light on) to 6 a.m. the following day (24 h fast). Resting metabolic rate (RMR) was calculated from the average of the 3 lowest points of energy expenditure values between 10 a.m. to 2 p.m. during the fasting period. Energy expenditure data was normalized to both lean mass and body weight. Locomotor activity was measured on x- and z-axes (horizontal and vertical activities, respectively) using infrared beams to count the number of beam breaks during the recording period.
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