Extended Data Fig. 1: Circadian and metabolic profiling in TRF-CR mice.

a, (left) Representative wheel-running behaviour and (middle) quantification of average daily onset of activity for TRF-Reg (n = 5) and TRF-CR (n = 5) mice throughout the duration of the 4-wk intervention (double plotted for clarity). (right) Background-corrected PER2::LUCIFERASE readings from excised suprachiasmatic nucleus (SCN) of TRF-Reg (n = 6) and TRF-CR (n = 6) mice. The grey shading indicates mean values ± SEM. Experiments were performed in 4–6 mo old male C57BL6/J mice. b, Body weight of TRF-Reg (n = 15) and TRF-CR (n = 15) mice over the duration of the 4-wk study. c, Blood glucose and serum insulin during oral glucose tolerance testing performed during the daytime (at ZT4) in TRF-Reg (n = 6) and TRF-CR (n = 6) mice. d, GAP/DHAP mass isotopomer distribution determined by mass spectrometry of liver from TRF-Reg (n = 3) and TRF-CR (n = 2) mice 30 min following i.p. administration of a 10:1 mix of U¹³C-lactate:U¹³C-pyruvate (1 g/kg). e-f, (left) Relative concentration of (e) NADH and (f) NAD⁺ by HPLC in liver of TRF-CR compared to TRF-Reg mice during the day (ZT4) (n = 12 for TRF-Reg, n = 11 for TRF-CR,) and at night (ZT16) (n = 9 for each diet) for each time point. (right) Concentration of (e) NADH and (f) NAD⁺ in ad lib fed wild-type mouse liver by HPLC every 6 hrs for 24 hrs (n = 7). g-h, Log₂-FC in daytime liver acyl-carnitine levels in (g) TRF-CR compared to TRF-Reg mice and (h) LbNOX- compared to null-overexpressing TRF-CR mice (n = 3). Data are presented as mean values ± SEM. Statistics were performed with unpaired, two-tailed student’s t test except as otherwise noted in the figure. *p < 0.5, ***p < 0.01.