Inhibition of nucleotide salvage constraints CVB3 replication.

A CVB3 luciferase reporter virus carrying a Renilla luciferase (Rluc CVB3) was used to study the replication of CVB3 in the presence or absence of compounds inhibiting the salvage pathways. A, E) Schematic representation of the purine (A) and pyrimidine (E) salvage pathway with the mode action of 6-MP (A) and CPU (E). B, F) Luciferase levels in cells infected with Rluc CVB3 (MOI 0.1) in the presence of different concentrations of 6-MP (B) or CPU (F). Cells were lysed at 2,4 or 6 hpi. Representative data of seven (B) or four (F) independent experiment are depicted (mean ± SD of 3 technical replicates). C, G) Percentage luciferase levels in 6-MP treated and CVB3-infected cells compared to control. Data represent mean ± SEM of seven (C) or four (G) independent experiments. D, H) Metabolomic analysis of hypoxanthine (D) or uridine (H) levels in 6-MP (D) or CPU (H) treated cells. Statistical analysis was done using an ANOVA. For 6-MP, not every concentration was included in all seven independent experiments, but every concentration is measured in at least three independent experiments with three replicates. ***p < 0.001, ****p < 0.0001.