Fig. 4

TRF reshapes the rhythms of metabolites and genes implicated in glycolysis. A Heatmap showing reprogramming of energy metabolites in tumor tissues of mice inoculated with A549 tumor cells upon TRF versus control treatment. B Abundance of beta-d-fructose 6-phosphate in tumor tissues from the mouse xenograft model collected at different times (ZT1, 5, 9, 13, 17, 21) over 24 h (Control: n = 5–6 mice per timepoint; TRF: n = 4–5 mice per timepoint). ZT0 indicates 10 pm, which was the start of the TRF intervention during the experiment. C Correlation analysis between beta-d-fructose 6-phosphate level and tumor weight (n = 55 mice). D–I qRT–PCR detects mRNA expression of rate-limiting enzymes implicated in beta-d-fructose 6-phosphate metabolism including D HK1, E GPI, F FBP1, G PFKP, H ALDOA, and I PGK1, in tumor tissues at different times over 24 h (Control: n = 3–4 mice per timepoint; TRF: n = 3–4 mice per timepoint). J The abundance of metabolites and genes involved in energy metabolism between the TRF and control groups. Log₂-fold changes in genes and metabolites are color-coded; red represents an increase and blue represents a decrease upon TRF compared to control. Black dots represent metabolites not detected by MS. Only enzymes detected by qRT‒PCR are shown. K mRNA expression of GPI gene in A549 tumor cells in vitro was measured by qRT‒PCR at different times over 24 h (n = 3). ZT0 indicates the end of the two cycles of 24-h TRF intervention. Data were analyzed and visualized with circacompare or by Pearson correlation analysis. Error bars, when present, show the SEM