Fig. 3. HIV-2 PR abolishes capped and uncapped mRNAs translation, but moderately inhibits EMCV IRES-driven translation. (A, B and C) A RRL under full translation conditions (see Materials and methods) was pre-incubated without (lanes 1) or with different amounts of HIV-2 PR (lanes 2–4: 3.5, 7 and 14 ng/µl). After 1 h at 30°C, Palinavir (10 µM final concentration) was added to the reactions. Different transcripts (schematically represented on the upper part of each panel) including (A) natural capped globin mRNA (2.5 ng/µl), (B) XL–EMCV mRNA (10 ng/µl) and (C) DC–HCV mRNA (10 ng/µl) were translated under these conditions. Samples were processed on 15% SDS–PAGE, submitted to autoradiography, the relative intensities of the bands were quantified and the results are presented at the bottom of each panel. (D) Aliquots of the samples 1–4 from (A) were resolved on 10% SDS–PAGE, proteins transferred to PVDF and the membranes were incubated with antibodies specific to the C-terminal part of eIF4GI (serum E). The resulting fragments and molecular weight markers (in kDa) are indicated on the figure.