FIG 4.

Small deletions in the minimal FXR-binding motif have deleterious effects on VEEV replication. (A) Schematic presentation of the VEEV/minRep/GFP genome, its HVD, and deletions introduced into the minRep sequence. The black box indicates a mutated HVD derived from artHVD. The red box indicates the position of the minimized element of the repeat. Dashed lines indicate identical amino acids. Gray boxes indicate positions of the deletions. (B) BHK-21 cells were electroporated with 3 μg of the in vitro-synthesized RNAs, and these cells were used to assess RNA infectivity in an ICA (see Materials and Methods for details). (C) The rest of the electroporated cells were seeded into 100-mm dishes. Media were harvested at 24 h postelectroporation (post EP), and viral titers were determined by a plaque assay on BHK-21 cells. The experiments were performed 2 times and demonstrated very similar differences. Significances of differences among the values were determined by one-way analysis of variance (ANOVA), followed by Dunnett’s multiple-comparison test (n.s., not significant; **, P < 0.01; ****, P < 0.0001). Data are presented as means with standard deviations (SD) (n = 3).