Fig. 1: Redox and metabolic rhythms in human red blood cells (RBCs).

A Experimental scheme for untargeted Human RBCs circadian sampling. RBCs from n = 3–4 human subjects were incubated with 11 mM glucose and kept under constant conditions (37 °C in continuous darkness) and sampling was performed every 3 h. Untargeted metabolite analysis was performed using GC-MS and LC-MS. B Heatmap of all cycling features ordered by the phase of the oscillation. Features extracted from spectra using Progenesis QI (LC-MS) and MassHunter (GC-MS) were used for the rhythmicity analysis using RAIN algorithm (*P < 0.05, FDR = 0.2). Data are Z-score normalized log10 intensities. C Principal components analysis (PCA) of the cycling metabolome. The two major components separate the data into samples corresponding to the circadian day (left, orange) and night (right, green) time-points. D Pathway analysis performed using Metabolite Set Enrichment Analysis (MSEA) using only identified 24-h rhythmic metabolites. The color bar represents the significance level, as shown in the table. *FDR < 0.01. E Table representing the enriched metabolic pathways with the corresponding metabolite hits, P values and FDR.