The effect of inhibition of AMPD2 and GMPS on cellular metabolism and 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, C) Luciferase levels in cells infected with Rluc CVB3 (MOI 0.1) in the presence of different concentrations of Decoyinine (A) or AMPD2 inhibitor (C). Cells were lysed at 2, 4, 6, 8 hpi. Representative data of three independent experiment are depicted (mean ± SD of 3 technical replicates). B,D) MTS assay performed in parallel with the luciferase assay depicted in A and C (mean ± SD). The cells were exposed to the different Decoyinine (B) or AMPD2 inhibitor (D) concentrations for either 8h after which a MTS assay was used to determine the viability of the cells. E) ¹³C-glucose isotope tracing study in control- and decoyinine (100 μM) or AMPD2 inhibitor (100 μM) treated HeLa R19 cells (three replicates; one independent experiment). The cells were lysed after 6h and measured by LC-MS to identify metabolites and quantify the different isotopologues. The different isotopologues are not distinguished in this Figure. Heatmap showing log2 fold changes of intracellular metabolites compared to control-treated cells.

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