Detailed Protocols for Mouse Aging Study

All the mouse samples were prepared at the University of Michigan in a specific pathogen-free colony kindly supplied by Dr. R. Miller. Genetic background and husbandry conditions for the Snell dwarf and GHRKO mice were described previously^{19 (link)}. Mice used for Rapamycin (encapsulated, used at 14 ppm), Canagliflozin (180 ppm), 17α-estradiol (17aE2) (14.4 ppm), Acarbose (1000 ppm), and Calorie-Restricted samples were of the UM-HET3 stock, produced as the offspring of CByB6F1/J mothers and C3D2F1/J fathers, as described in this article^{20 (link)}. The base diet was Purina 5LG6. To monitor specific-pathogen status, sentinel mice were exposed to spent bedding for two weeks prior to testing and all tests were negative for the entire aging colony during the experimental period. The protocols were reviewed and approved by the University of Michigan’s Institutional Animal Care and Use Committee. The metadata of the mouse liver, kidney, gastrocnemius muscle tissues, and plasma samples are provided in Supplementary Table 1.