Figure 2. Respiratory and glycolytic flux analysis of HCV-infected and sofosbuvir-treated primary human hepatocytes.
(a) Metabolic flux balance map of JFH-1-infected primary hepatocytes compared to naive cells. “Upregulated flux” and “downregulated flux” refer to changes in after infection. Statistically significant changes determined by P < 0.05 (n = 3). Metabolic fluxes measured by targeted metabolomics or HPLC are marked with an asterisk. TCA, tricarboxylic acid. (b) Flux measurements (Supplementary Fig. 2) of mitochondrial basal respiration, oxidative phosphorylation, glycolysis and fatty acid oxidation, normalized to naive cells. JFH-1 infection caused a 34% drop in mitochondrial function and a 36% drop in oxidative phosphorylation (P < 0.0001, n = 3). Sofosbuvir reversed this effect, but function remained significantly lower in sofosbuvir-treated cells than in naive cells (P < 0.001, n = 3). JFH-1 infection increased fatty acid oxidation by 85% (P < 0.02, n = 3) and glycolysis by 169% (P < 4 × 10−5, n = 3); both effects were drastically diminished by treatment with sofosbuvir (P > 0.05, n = 3). (c) Metabolic phenotype graph of naive cells, JFH-1-infected cells and infected cells treated with sofosbuvir, showing the metabolic shift toward glycolysis in HCV-infected cells and its reversal after sofosbuvir treatment (n = 3). **P < 0.001; n refers to experimental replicates.