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Fig. 7. Inhibition of translation by HOPS in vitro and reduction of proliferation in vivo. (A) Luciferase cDNA was translated in an in vitro translation system with and without (C: control) different concentrations of purified recombinant HOPS protein (60-420 nM). Similar experiments were performed with or without purified recombinant GST protein. eEF-1A expression was analyzed by western blotting on reticulate lysates with different HOPS concentrations. (B) The synthesized luciferase was quantified by densitometric analysis after gel electrophoresis. The average results from three experiments are shown (in arbitrary units). Black bars: the amount of luciferase after the addition of different concentrations of HOPS; gray bars: the amount of luciferase after the addition of different concentrations of GST. (C) Percentage thymidine incorporation by stable H-35 stable clones (C1-C3) overexpressing HOPS with respect to H-35 control. (D) Number of puro selected NIH-3T3 cells at different days after infection. The gray line shows the number of control cells infected with plasmid Puro; the black line shows the number of cells infected with plasmid Puro containing HOPS.
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