A. Schematic depiction of selection procedure. The single virion represents a selected clonal, plaque-purified virus; whereas the multiple virions represent diverse virus populations (quasispecies). B. Plaque assay analysis of the fold increase in titer for EHDV-TAU, compared to EHDV2-IBA, in LNCaP cells. Panel depicts typical images of the plaque assays; the dilution employed appears above the respective wells. Left inoculum (10−3 dilution); right, dilutions (10−3 and 10−10) of the virions (of the indicated viruses) produced in LNCaP cells (60 h infection) C. Immunoblot analysis of NS3 production and STAT1 phosphorylation in EHDV-TAU-infected cells. Lysates (100 μg protein) of DU145 or LNCaP cells, infected or not with EHDV-TAU (0.05 pfu/cell, 45 h) were separated by SDS-PAGE, blotted and probed with antibodies against the indicated proteins. α-tubulin was used as a loading control. D-E Exogenous addition of IFNα blocks EHDV-TAU infection in DU145 cells, but not in LNCaP cells. D. Panels depict typical fields of DU145 and LNCaP cells, stained for DAPI (blue, left panels) and NS3 (red, right panels) under the indicated conditions: uninfected, EHDV-TAU-infected (45 hpi, 0.05 pfu/cell), or EHDV-TAU infected (45 hpi, 0.05 pfu/cell) treated with IFNα (200U/ml, 45 h). Bar, 100 μm. E. Graph depicts the mean ± SE percentage of NS3 positive cells. Quantification of percentage NS3 positive cells was from multiple (n=5) randomly selected fields, imaged under the same conditions as in (D). *, p<0.05.