Fig. 1.

Inhibition of cap-dependent translation by 4E2RCat. (A) Schematic diagram illustrating the structure of 4E2RCat. An 8-point dose-response curve of 4E2RCat in a TR-FRET assay is provided to the right. (B) Inhibition of translation by 4E2RCat. Schematic representation of FF/HCV/Ren bicistronic construct used for in vitro translation studies (top). In vitro translations were performed in Krebs extracts programmed with FF/HCV/Ren in the presence of [³⁵S]methionine, and a representative autoradiograph of the products after fractionation on 10% SDS-PAGE is provided (bottom left). Translations contained vehicle (1% DMSO) (lane 1), 500 μM m⁷GDP (lane 2), 500 μM GDP (lane 3), 50 μM anisomycin (lane 4), the indicated concentrations of 4E2RCat (lanes 5 to 10), or no RNA (lane 11). FF and Ren RLU values (relative to DMSO controls) from two independent experiments are provided with the standard errors of the means (SEM) indicated (bottom right). (C) Schematic representation of FF/EMCV/Ren bicistronic construct used for in vitro translation studies (top). RLU values (relative to those of the DMSO control) from two independent in vitro translations performed in Krebs extract programmed with FF/EMCV/Ren mRNA are provided with the SEM indicated (bottom).