An osmotic pump (Alzet) with a 200 μL reservoir volume and a 2-week release duration was filled with 75 μg of recombinant BMP3B (R&D Systems) dissolved in 4 mM HCl. The control pump was filled with 4 mM HCl. Filled pumps were subcutaneously transplanted into 26-month-old C57BL/6N females. After 2 weeks, the mice were housed in a metabolic cage equipped with a running wheel (Shinfactory) for 3 days. Mice were acclimated for the first 24 hours and then monitored for 48 hours. The oxygen consumption, carbon dioxide production, and energy expenditure were measured using an Oxymax open-circuit indirect calorimetry system (Columbus Instruments). Home-cage activity and wheel-running activity were measured using the Actimo dual activity monitoring system (Shinfactory). Mice were subjected to the grip strength test prior to euthanizing.

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