aniPOND detects viral and cellular proteins associated with replicated viral genomes.

<p>(A) ICP4 was detected in protein eluates from aniPOND experiments carried out on wild type KOS or UL2/UL50 mutant virus by western blotting. AniPOND was carried out on virus grown in the presence (+) or absence (-) of 2.5 μM EdU at 4–8 hpi and/or 33 μM acycloguanosine (ACG) throughout infection. (B) DNA eluted from streptavidin-coated beads during aniPOND experiments in (A) is viral. The amount of viral DNA present in cell lysates (input) and eluted from beads (bound) during aniPOND experiments was measured by qPCR of the viral TK gene. The ratio of viral DNA (vDNA) to total DNA was calculated and is presented in log %vDNA. Values for virus grown in the presence of ACG are too small to be displayed on this graph. These experiments were carried out with the UL2/UL50 mutant virus. (C) Pie charts summarize proteins that were identified by mass spectrometry of protein eluates from aniPOND carried out on the UL2/UL50 mutant grown in the presence of EdU at 4–8 hpi. Pie charts represent proteins that were identified with high confidence in independent duplicate experiments. Values indicate the number of proteins identified for each functional category. (D) Venn diagrams depict the overlap of proteins identified by iPOND and aniPOND carried out on the UL2/UL50 mutant 8 hpi.