(A) NAD+ levels in metabolic tissues between young and old mice. Pancreas, liver, WAT, and skeletal muscle were collected from young (n=5–11) and old (n=5–15) mice at 3–6 and 25–31 months of age, respectively. (B) Glucose tolerance in aged, naturally occurring diabetic male mice before (closed circles) and after (open circles) a single dose of NMN (500 mg/kg body weight) (n=11). The areas under each glucose tolerance curve are presented next to the glucose tolerance curves. (C) Plasma insulin levels were measured during IPGTTs (n=5). (D and E) Glucose tolerance (D) and lipid levels (E) in aged HFD female mice before (closed circles) and after (open circles) NMN (n=5). Fasted plasma samples were collected from the same mice and subjected to the measurements of cholesterol (Chol), triglycerides (TG), and non-esterified free fatty acids (FFA). Data were analyzed by Student’s unpaired t test (A), paired t test (B–D), and one-way ANOVA with the Fisher’s PLSD post-hoc test (E). All values are presented as mean ± SEM. *P < 0.05; **P <0.01; ***P < 0.001.