FIG 1.

Generation and characterization of rLCMV/GFP-P2A-NP. (A) Schematic of rLCMV/GFP-P2A-NP. The rLCMV/GFP-P2A-NP was generated as described in Materials and Methods. (B) Comparison of rLCMV/GFP-P2A-NP and r3LCMV/GFP propagation in cultured cells. Vero cells were infected with either the rLCMV/GFP-P2A-NP or r3LCMV/GFP at an MOI of 0.01 or 0.1; at 36 h p.i., the cells were fixed with 4% PFA, and GFP expression was assessed by fluorescence microscopy. The panels show representative fields using the same magnifications and exposure times. (C) Comparison of rLCMV/GFP-P2A-NP and rLCMV/WT growth kinetics in cultured cells. BHK-21, A549, and Vero cells were infected with rLCMV/WT or rLCMV/GFP-P2A-NP at an MOI of 0.01 or 0.1. At the indicated times, TCS were collected, and virus titers were determined. The data represent the means ± the standard deviations (SD) of two independent experiments. Each independent experiment consisted of three replicates. (D) Inhibition of rLCMV/GFP-P2A-NP by Rib. Cells were treated with Rib at the indicated concentrations and infected with rLCMV/GFP-P2A-NP (MOI = 0.01). At 36 h p.i., the cells were fixed with 4% PFA, and GFP expression was examined by fluorescence microscopy. Nuclei were visualized by DAPI staining. The limit of detection (LoD; shown as dashed lines) indicates the threshold of level of detection of infectious virus.