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Figure 3 | Virology Journal

Figure 3

From: Origin-independent plasmid replication occurs in vaccinia virus cytoplasmic factories and requires all five known poxvirus replication factors

Figure 3

Intracellular localization of replicated plasmid containing tandem lac O repeats by staining with an MPB-lac I fusion protein. HeLa cells were transfected with pSV2-dhfr-8.32 containing lac O tandem repeats and 24 h later were infected with 3 PFU per cell of vV5D4 expressing V5-tagged uracil DNA glycosylase. At 12 h after infection, cells were fixed, permeabilized, incubated with MBP-lac I and rabbit antibody to MBP (anti-MBP) and mouse monoclonal antibody to the V5 epitope (anti-V5) followed by Cy5-conjugated donkey anti-mouse IgG and Texas red dye conjugated donkey anti-rabbit IgG. Cells were counterstained with Hoechst dye and analyzed by confocal microscopy. Colors: deep blue, Hoechst dye; red, Texas red; white, Cy5; light blue, overlap of Texas red and Cy5; yellow, overlap of Hoechst and Cy5.

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Virology Journal

ISSN: 1743-422X