Fig 5. NxN1/2 mutation in ICP0 results in delayed and reduced vital DNA and protein synthesis.

(A) U2OS cells, and (B) HepaRG cells were infected with HSV-2-ICP0-GFP (ICP0-GFP) or HSV-2-ICP0^NxN1/2-GFP (NxN1/2-GFP) at an MOI of 2 or 1 pfu/cell, respectively. Infected cells were harvested at the indicated time points post infection (p.i.) for isolation of genomic and viral DNA. Viral DNA copies were determined by quantitative real-time PCR using oligonucleotide-primers and probes directed against the UL27 gene encoding glycoprotein gB. Copy numbers were normalized to the sequence of the human single-copy beta-globin gene (HBG). (C) U2OS cells, and (D) HepaRG cells were infected with HSV-2-ICP0-GFP or HSV-2-ICP0^NxN1/2-GFP at an MOI of 2 for up to 24 hours, or 1 pfu/cell for 12 hours, respectively. At the indicated time points, cells were harvested. Whole cell lysates were analyzed by SDS-PAGE and Western blot against the indicated proteins. Signals were quantified in S2 Fig.