FIG. 6.

Expression levels of standard and truncated H proteins analyzed by cell surface biotinylation assay, Western blot analysis, and metabolic labeling. Vero cells were transfected with plasmids carrying the H genes indicated. At 24 h posttransfection, the cell surfaces were labeled with biotin and lysed. After immunoprecipitation of the H proteins, the samples were divided into two parts. Each was subjected to SDS-PAGE and blotted to nitrocellulose. (A) One blot was incubated with streptavidin-peroxidase to visualize surface-labeled H proteins. (B) The second blot was stained with a polyclonal MV antiserum and peroxidase-conjugated anti-rabbit immunoglobulins to detect the total amount of H proteins. (C) Transfected cells were radiolabeled with [³⁵S]methionine and [³⁵S]cysteine for 10 or 30 min, respectively. After lysis, H was immunoprecipitated and separated on a 10% SDS gel. (D) For pulse-chase analysis, cells were radiolabeled for 45 min and then incubated in chase medium for 0, 90, or 180 min. Quantification of the signals was performed using a BioImager.