Figure 4. 6AP and GA do not affect the translational activity of the ribosome both in vivo and in vitro.

a. The effect of the indicated compounds on general in vivo translation in living yeast cells was evaluated. Briefly, various drugs or DMSO (the compounds vehicle) alone were added to yeast cells in exponential growth in YPD rich medium at a final concentration of 100 µM. After 10 minutes, radiolabelled [³⁵S] methionine was added for 10 minutes. The cells were harvested to prepare cell extracts which were then analyzed by SDS-PAGE followed by autoradiography. Note that, with the exception of cycloheximide (CHX, a known inhibitor of general translation), none of the tested compounds has a significant effect on global protein synthesis. b. WT yeast cells were grown in the presence of DMSO (control left panel) or 100 µM 6AP (this concentration allows an efficient cure of prion) and cell extracts were then analyzed by 2D-SDS-PAGE followed by autoradiography. Note that no significant difference was observed between 6AP-treated cells and control cells. c. Effect of the indicated compounds on in vitro expression of EF1A using a commercial kit (Ambion). Note that only cycloheximide (CHX) significantly inhibits synthesis of EF1A.