Fig. 6: Metabolic inhibition abolishes rhythmic redox cofactor accumulation in human red blood cells (RBCs).

A Schematic showing experimental protocol used to collect samples. RBCs from n = 3–4 human subjects were incubated with 11 mM 1, 2-13C2-glucose and kept under constant conditions (37 °C in continuous darkness) and sampling was performed every 4 h. Cells were treated with metabolic inhibitors, HA and 6AN at the starting of the experiment. B Effect of metabolic inhibitors HA and 6AN on the rhythmicity of the redox coenzyme, NADH. Graph bars present mean ± s.e.m (n = 6 biological replicates) C Effect of metabolic inhibitors HA and 6AN on the rhythmicity of the redox coenzyme, NADPH. Graph bars present mean ± s.e.m (n = 6 biological replicates). D Schematic showing experimental protocol used to collect samples after treating with inhibitors Conoidin A and MG-132. E RBCs were treated with inhibitors of PRDX oxidation (Conoidin A) or PRDX oxidation rhythms (MG-132) and their effect on glucose flux was assessed. All data are mean ± s.e.m. (n = 4 biological replicates). The 24-h rhythmicity P-value (determined by RAIN) for each profile is shown with each plot, in addition to the best fit period of the rhythm if this was not 24 h. ns, not significant (P > 0.05).