Fig 1.

HSV-1 lacking VP22 fails to package VHS. (A) Fifty PFU of s17 (Wt) and Δ22 viruses were plated on Vero cells and fixed and stained with crystal violet 4 days later. (B) The relative areas of Wt and Δ22 plaques from 3 separate experiments was measured using Image J software. Values are shown as means ± standard errors of the means of 10 plaques. (C) HeLa cells treated with 5 μg/ml actinomycin D were infected with s17 (Wt), Δvhs, or Δ22 viruses or left uninfected (M). Five hours later the cells were metabolically labeled with 50 μCi/ml [³⁵S]methionine for 1 h, and total lysates were analyzed by SDS-PAGE and autoradiography. (D) Gradient-purified virions from s17 (Wt) and Δ22 viruses were analyzed by SDS-PAGE, followed by Coomassie blue staining (left) or Western blotting (right) with antibodies against the major capsid protein VP5 and the tegument proteins VP22, VP16, and VHS.