Figure 4.

(A) Uncapped polyadenylated (−/+) and uncapped non-polyadenylated (−/−) EMCV-renilla RNAs were translated at 0.5 mg/ml during 30 min at 30°C in both the nuclease-treated (left panel) and untreated RRLs (right panel) that were pre-incubated with 0, 0.2, 0.4 or 0.6 µl of an in vitro-synthesized FMDV L protease (see Materials and Methods section). Translation products were resolved by 15% SDS-PAGE followed by autoradiography. Densitometric analysis of the renilla protein product was performed and expressed as percentage of the control (100%, no l protease added) at the bottom of each panel. (B) At the end of the in vitro translation incubation, samples (1 µl) from the experiment described above were run on a 10% SDS–PAGE and the proteins transferred to PVDF membrane and incubated with antibodies specific to the C-terminal part of eIF4GI.