Fig. 2

Bioenergetics of WNV infection in cultured cells. A Changes in mitochondrial respiration profile induced by WNV replication. Real-time monitorization of oxygen consumption rate (OCR) in Vero cells infected or not with WNV RVPs at 1 or 48 hpi (n = 5). Arrows indicate the time of injection of oligomycin, FCCP and rotenone/antimycin A. Respiratory parameters determined are indicated by colored areas. B Comparison of respiratory parameters determined by OCR measurement between control (uninfected) and infected cells with RVPs at 48 hpi. Fold change over uninfected at 1 hpi is represented. ***, P < 0.001 for two-way ANOVA and Sidak’s multiple comparisons test (n = 9). C Changes in glycolytic rate induced by WNV replication. Real-time monitorization of extracellular acidification rate (ECAR) was measured in Vero cells infected or not with WNV RVPs at 1 or 48 h pi (n = 6). Arrows indicate the time of injection of glucose, oligomycin and 2-DG. Glycolytic parameters determined are indicated by colored areas. D Comparison of glycolytic parameters determined by ECAR measurement between control (uninfected) and cell infected with SRIPs at 48 hpi. Fold change over uninfected samples at 1 hpi is represented. *P < 0.05; **P < 0.01; ****P < 0.0001 for two-way ANOVA and Sidak’s multiple comparisons test (n = 10). E–H Effect of glucose metabolism manipulation on WNV infection. Vero cells were infected with WNV (MOI of 1 PFU/cell) and subjected to glucose depletion (E), treatment with 10 µM AL-429 (F), 10 mM 2-DG (G) or 50 mM oxamate (H) and virus yield was determined at 24 hpi by plaque assay. Cell viability was evaluated in uninfected cells treated in parallel by quantification of cellular ATP. **P < 0.01, ***P < 0.001; ****P < 0.0001 for unpaired t-test applying Welch’s correction when differences between variances were found (n = 4–6)