FIG 7.

Translation promoted by the DENV 5′ UTR is activated for the proteolytic cleavage of eIF4G by HRV 2A protease. The dl HCV IRES or dl 5′UTR DENV plasmid (200 ng) was transfected into HEK 293T cells together with a plasmid expressing either wild-type HRV 2A protease (p2A-wt) or an inactive mutant (p2A-mut) (500 ng). (A) The cleavage of eIF4G was monitored by Western blotting using polyclonal antibodies against eIF4GI. The eIF4G cleavage product (CP) has been characterized previously (34). (B and C) RLuc (B) and FLuc (C) activities were measured and are expressed in relative light units (RLU). Statistical analysis was performed with a two-tailed t test. Asterisks indicate P values of <0.05; ns, no significant difference. Values represent means (± standard errors of the means) for three independent experiments, each conducted in duplicate.