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For protein normalisation, protein concentration was measured using the Bradford method with the Protein Assay CBB Clean Up Kit and Protein Assay CBB Solution (both from Nacalai Tesque Inc.), with the standard calibration curve prepared using bovine serum albumin solution (Nacalai Tesque Inc.)

Samples were cleared by centrifugation at 4 C 10,000 rpm for 5 min, and the supernatants were analyzed and quantified with the SAM standard curve

Recently, for the first time, the safety of single-day NMN oral administration was reported in humans 20

Reactive oxygen species (Ros-Induced) Ros release: a new phenomenon accompanying induction of the mitochondrial permeability transition in cardiac myocytes