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. 2008 Mar 5;82(10):5054–5067. doi: 10.1128/JVI.02174-07

FIG. 1.

FIG. 1.

UL97 reduces the number of PML domains in COS7 cells by a kinase-dependent mechanism. Cells were transfected with plasmids expressing ppUL44, IE1, pp71-V5, UL97-V5, UL97-V5 K355M, and UL97-V5, with the addition of MBV, as indicated in the figure. PML domains were visualized with a rabbit polyclonal antibody to SP100 and a FITC-conjugated secondary antibody (green staining), and examples of these sites are indicated by arrows. The expression of viral proteins was confirmed by staining with monoclonal antibodies labeled with Texas Red (red staining). All nuclei were stained with DAPI (4′,6′-diamidino-2-phenylindole) (blue staining). Values shown in each panel are the average numbers of PML domains in cells expressing the viral proteins and reflect the average of at least 50 cells, with the standard deviations as shown. Control cells expressing ppUL44 contained an average of 4.7 PML domains (A), while positive control cells expressing IE1 contained significantly fewer PML domains (B). The expression of pp71 resulted in the recruitment of this protein to PML domains, but did not affect their number (C). Cells expressing UL97 also contained reduced numbers of PML domains (D) and was dependent on its kinase activity, since neither the K355M mutant nor UL97 in the presence of MBV significantly reduced their numbers (E and F).