FIG. 6.

Effects of NEM on the UL16-capsid interaction. (A) Extracellular virions were treated with or without NEM at 37°C. Viral membranes were removed with NP-40 treatment, and the capsids were pelleted through a 30% sucrose cushion prior to immunoblot analyses with antibodies specific for VP5, VP16, and UL16. (B) Extracellular virions were treated with or without NEM for 30 min and then treated with or without NP-40. The released capsids or intact virions (C or V, respectively) were pelleted through a 30% sucrose cushion prior to immunoblotting with the indicated antiserum. (C) Virions present in the medium were concentrated by centrifugation through a sucrose cushion and resuspended in a small volume of TNE buffer. The sample was divided into two equal portions, and these were treated with or without NEM for 30 min. Both samples then received NP-40 for 15 min at room temperature prior to sedimentation through parallel 20 to 50% sucrose gradients. Fractions were collected from the top and analyzed by immunoblotting with antibodies specific for VP5 and UL16. (D) Gradient-purified capsids treated with NEM or carrier prior to lysis for 15 min at 37°C from infected cytoplasm and medium were analyzed by immunoblotting for levels of VP5 and UL16. Gradients were run in duplicate. α, anti.