UGGT1 enhances 3D viral polymerase levels in a viral protein 3A-associated membrane fraction.

<p>(A) RD cells were co-transfected with pFLAG-3A and pFLAG-3D; pFLAG-3AB and pFLAG-3D; or pFLAG-3D only, and harvested at 48 h post-transfection. Membrane fractions were isolated and subjected to Western blot analysis. (B) Cells were transfected with pFLAG-3A, pFLAG-3AB, and pFLAG-vector and harvested at 48 h post-transfection. Membrane fractions were separated and subjected to Western blot analysis. (C) Cells were transfected with NC or UGGT1 siRNA for 48 h, and then co-transfected with pFLAG-3A and pFLAG-3D, or pFLAG-3AB and pFLAG-3D. Cells were harvested at 48 h post-transfection, and the membrane fractions were extracted and subjected to Western blot analysis. Anti-UGGT1, anti-3D and anti-3A antibodies were used. The same blot was probed with anti-CNX and anti-HSP90 antibodies. (D) Cells were transfected with NC or UGGT1 siRNA for 48 h, and then co-transfected with pFLAG-3D. Cells were harvested at 48 h post-transfection, and the membrane fractions were extracted and subjected to Western blot analysis. Anti-3D, anti-UGGT1 and anti-CNX antibodies were used. Results are representative of at least three independent experiments.