Figure 7.

N replaces eIF4G. (A) HeLa cells were co-transfected with a plasmid expressing reporter GFP, along with increasing amounts of pF/HRV-16 2A, which expresses the 2A protease of HRV-16. GFP expression was quantified using flow cytometry as in Figure 1 (dark bars) and GFP mRNA was quantified using real-time PCR (light bars). (B) Cells were transfected with a constant amount of GFP expression plasmid, a constant amount of 2A expression plasmid (0.05 μg) sufficient for significantly reducing translation of the reporter gene, and increasing amounts of an N expression plasmid. GFP expression (dark bars) and steady-state GFP mRNA (light bars) were quantified as in (A). In the experiments of (A, B), the total amount of DNA used in the transfections was held constant by the addition of parental vector.