PRF disrupts equimolarity of viral protein synthesis in replicon-transfected and virus-infected cells.

<p>(<b>A</b>) KUNRep-WT and KUNRep-A30A′ replicons were electroporated into BHK cells and 48, 72, 96 hpe lysates were harvested and total proteins were separated by SDS-PAGE and detected by Western blot with anti-β-gal and anti-NS3 or (<b>B</b>) anti-β-gal and anti-NS5 antibodies. Ratios of β-gal/NS proteins were calculated and plotted as a function of KUNRep-WT ratios. (<b>C</b>) BHK cells were infected with WNV_KUN WT or WNV_KUN A30A′ virus and at 48 and 72 hpi proteins were pulse-labelled with ³⁵S methionine/cysteine with and without brefedrin A (BFA) treatment, harvested with RIPA buffer and immuno-precipitated with anti-E and anti-NS5 antibodies. Precipitated proteins were separated by SDS-PAGE and visualised by exposure to X-ray film. Ratios of E/NS5 were calculated as above and represent 2 independent experiments. Error bars represent standard error of the mean of 2 independent experiments.