Virion Assembly Factories in the Nucleus of Polyomavirus-Infected Cells
Figure 5
PyV DNA and T-antigen are localized near PML-NBs.
C57 MEFs were infected at an MOI of 30–40 pfu/cell. At 24 or 28 hpi cells were fixed, permeabilized, and co-stained with either anti-PML, anti-Tag and/or anti-MRE11 antibodies followed by AlexaFluor-conjugated secondary antibodies, a fluorescently-labeled PyV DNA FISH probe, and DAPI staining of nuclei. A) PyV-infected C57 MEFs at 24 hpi stained for either PML (top) or Tag (bottom) followed by fluorescent in situ hybridization (FISH) for PyV DNA. B) PyV-infected C57 MEFs at 28 hpi co-stained for PML and Tag. C) C57 MEFs at 24 hpi co-stained for either MRE11 and PyV DNA (by FISH) or MRE11 and Tag. All images represent a 0.1 mm z-stack slice. Insets show enlarged regions from image to illustrate the localization of proteins in relation to each other (B) or PyV DNA (C).