FIG. 7.
HRV2 is converted to C-antigenic particles but lysosomal degradation is prevented in nocodazole-treated cells. (A and B) HeLa cells were preincubated (with or without 20 μM nocodazole) for 30 min at 34°C in infection medium and infected with [35S]HRV2 (105 cpm) at 34°C. Aliquots were removed at 5, 10, 20, and 30 min. Samples were rapidly cooled, cells were pelleted, plasma membrane bound virus was removed with PBS–10 mM EDTA, and cell pellets were lysed in RIPA buffer. Lysates (A) and cell supernatants (B) were subjected to immunoprecipitation with monoclonal antibody 2G2, which is specific for C-antigenic particles, and polyclonal antiserum. (C) Cells were preincubated and infected as in panel A for 30 min. Unbound virus was removed with PBS, and incubation was continued with or without nocodazol for the times indicated. Virus was recovered from cell lysates by immunoprecipitation with 2G2. Viral proteins were analyzed by SDS-gel electrophoresis followed by autoradiography.