The inhibitory effect of MG132 on HEV replication does not result from the inhibition of translation. (A) Effect of MG132 treatment on GFP synthesis. A subclone of Huh7 cells was transfected with a similar amount of pAcGFP N1 plasmid in six-well plates. Inhibitor treatment started 1 day posttransfection, cells were harvested on the fifth day posttransfection, and immunoblotting was performed with anti-GFP polyclonal serum produced in a rabbit. Lane 1, mock-transfected cells; lane 2, pAcGFP N1-transfected cells; lane 3, pAcGFP N1-transfected cells with 1 μM MG132 treatment. (B) Effect of MG132 treatment on ORF2 protein synthesis. A subclone of Huh7 cells were transfected with a similar amount of pTrix-neo-ORF2 (Δ1–111) plasmid in six-well plates. Inhibitor treatment started 1 day posttransfection, cells were harvested on the fifth day posttransfection, and immunoblotting was performed with chimpanzee 1313 anti-HEV immune serum. Lane 1, mock-transfected cells; lane 2, pTrix-neo-ORF2 (Δ1–111)-transfected cells; lane 3, pTrix-neo-ORF2 (Δ1–111)-transfected cells treated with 1 μM MG132.