Figure 2

ORF6 expression in Vero E6 cells does not cause a reduction in total cellular protein. Vero E6 cells were transfected for 16 hours with increasing amounts of pXJ3'-ORF6 or an empty vector, pXJ3'HA. Cells were starved of methionine and cysteine for 1 hour and labeled with a mixture of 35S-labelled methionine and cysteine for 30 minutes, then subjected to SDS-PAGE separation and autoradiography (lower panel). Quantification of the signals in each lane of the resulting autoradiograph was performed and the readings, after normalization to the reading in the lane from the untransfected cells, are shown at the bottom of the autoradiograph. A fraction of lysate was subjected to Western Blotting using an antibody directed against the ORF6 protein (upper panel). Similar results were obtained in two independent experiments, and a representative set of data is presented.