Biochemical correlates of changes in RBC oxygen-handling function. (A) Metabolomics of RBC lysates. Heatmap shows 130 metabolites significantly affected by hypoxic storage or rejuvenation (2-way ANOVA: time and treatment with P < .05). (B) Scatter plot shows effect of hypoxic storage or rejuvenation for the differentially abundant metabolites. Green symbols show concordantly responding metabolites. Black symbols show metabolites that respond oppositely to hypoxia and rejuvenation. Red symbols show metabolites affected by hypoxic storage only. Blue symbols show metabolites affected by rejuvenation only. Paired t-test, P < .05. Inset highlights the 3 chemical components of rejuvenation solution. (C) Pearson’s correlation coefficient between specific metabolite and time constant (τ; top) or SSC (bottom). Color coding refers to scheme described in panel E. (D) Top 3 negative correlations to time constant (top) or SSC (bottom). Bars show time constant or SSC data from Figure 2; circles show metabolite levels. Number in italics refers to correlation coefficient. (E) Analysis for the top 3 positive correlations. (F) Multivariate regression analysis using the top 3 negative and top 3 positive correlates. For time constant τ: R2 = 0.6482, F = 10.75, P < .0001. For SSC: R2 = 0.9128, F = 71.53, P < .0001.