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. 2008 Jun 18;82(17):8362–8372. doi: 10.1128/JVI.00900-08

FIG. 5.

FIG. 5.

The C terminus of Nbs1 is required for inhibition of Ad replication. (A) Schematic of the domains in wild-type Nbs1 and Nbs1 truncation mutants. (B) Immunoblot of lysates from NBS cells expressing wild-type and mutant Nbs1 proteins. GAPDH served as a loading control. (C) The C terminus of Nbs1 is required for inhibiting mutant Ad DNA replication. NBS cells expressing wild-type or mutant Nbs1 were infected with the E4-deletion mutant virus (dl1004), and DNA was extracted over a time course of infection for analysis by qPCR. Accumulation of viral DNA is represented as the fold increase over input viral DNA, determined at the 4-h time point. (D) Localization of Nbs1 and Rad50 proteins in the absence or presence of virus infection. NBS-derived cell lines either uninfected or infected with E4-deleted virus dl1004 were fixed and analyzed by immunofluorescence at 24 hpi. Virus replication centers were visualized in infected cells by staining with an antibody to DBP. Cell nuclei were marked by staining with DAPI.