iPOND detects viral and cellular proteins associated with replicated HSV-1 genomes.

<p>(A) ICP4 was detected by western blot of protein eluates from iPOND carried out on viral genomes grown in the presence of EdU at 4–6, 6–8, and 8–12 hpi. The control was iPOND carried out on virus grown in the absence of EdU (−) and harvested 8 hpi. Purified ICP4 is shown. (B) DNA eluted from streptavidin-coated beads during iPOND experiments in (A) is viral. The amount of viral DNA present in cell lysates (input) and eluted from beads (bound) during iPOND experiments was measured by qRT-PCR of the viral thymidine kinase (TK) gene. The ratio of viral DNA (vDNA) to total DNA was calculated and is presented as log %vDNA. (C) Pie charts summarize proteins that were identified by mass spectrometry of protein eluates from iPOND carried out 6, 8, and 12 hpi with UL2/UL50 mutant virus. Values indicate the number of proteins identified for each functional category. (D) Venn diagrams depict the overlap of proteins identified by iPOND at each time point.