Skip to main content
. 2009 Feb 5;28(6):652–662. doi: 10.1038/emboj.2009.15

Figure 7.

Figure 7

E4orf3 prevents ATR-dependent damage signaling induced by nonviral sources. (A) U2OS cells transfected with a plasmid vector expressing Ad5-E4orf3 were laser microirradiated. Cells were fixed and stained for endogenous Nbs1 and Mdc1. Arrows indicate cells with E4orf3-induced tracks of Nbs1. (B) Stable U2OS cell lines expressing Nbs1–2GFP were transfected with a plasmid expressing Ad5-E4orf3, together with prRFP-C1, which was used as a marker for transfected cells. Cells were laser microirradiated (as indicated by dashed line) and images show the recruitment of Nbs1 to sites of damage. (C) HeLa cells were mock infected, or infected with E1-deleted recombinant Ads expressing GFP (rAd-GFP) or E4orf3 (rAd-E4orf3). At 24 hpi, the cells were mock treated or treated with 2 mM hydroxyurea (HU) for 2 h, and the cells were harvested for immunoblotting. Cellular proteins were detected with antibodies to RPA32 and specific phosphorylated sites at RPA-S4,8 and Nbs1-S343. Ku86 served as a loading control.