FIG. 2.
Effects of phallacidin treatment on EIAV production. (A) Daily EIAV production from phallacidin-treated (▪) and -untreated (□) EIAVuk-infected ED cells. Parallel cell cultures grown on a six-well plate were incubated with 25 μM phallacidin at 37°C for 20 h (indicated by the gray bar), at which time drug-free culture medium was added and replaced daily. Virion production during each 24-h interval was assayed by determining the RT activity of virions released into the culture medium. (B) EIAV production at 2-h intervals. Parallel cells were grown and treated as for panel A except that the culture medium used for RT assay was collected at 2-h intervals. (C) Western blot assay showing EIAV production at the indicated time points. EIAV particles pelleted from equal amounts of culture medium (see panel B) at the indicated time points were resolved through a 4 to 15% SDS-PAGE gel and probed by the reference serum. The intensity of the virus-specific capsid protein was used to quantify virion production. (D) EIAV production from infected equine macrophages at the maximal inhibition time (day 3). The data represented the mean values of at least three independent experiments done in duplicate.