FIG 2.

Effect of selected substrate supplementation on RV replication and Vero cell growth rate. d-Glucose (Glc), l-glutamine (Gln), and sodium pyruvate (Pyr) as important carbon sources for mammalian cells were used for selected substrate supplementation (suppl.) of mock- and RV-infected Vero cells. (A) Summary of the metabolic pathways fueled by the applied nutrients. AA, amino acid; α-KG, α-ketoglutarate. (B) Schematic illustration of the application of the indicated supplements either 4 h after plating (supplementation before infection) or 2 hpi (supplementation after infection). (C) Viral titer (assessed for the Therien strain by a standard plaque assay) was determined for the indicated substrate supplementation conditions at 24, 48, and 72 hpi. CTL, control in maintenance medium. (D) For assessment of cell morphology and density under given substrate supplementation conditions, phase-contrast images of mock-infected Vero cells were obtained 24 h after plating before the initiation of infection with RV. (E) The number of live and dead cells was determined by trypan blue exclusion assay at 24 and 48 h after cell plating for the indicated substrate supplementation conditions. (F) Viral titers were determined by plaque assay (Therien and Wb-12) and focus-forming assay (03-03703 and 07-00426) at 24 hpi.