Figure 3.

Redistribution of UBF and Fibrillarin depends on viral DNA replication. BAC16-BFP-mCherry-infected iSLK cells were treated for 48 h with 1µg/mL Dox and 1 mM n-Butyrate to induce lytic reactivation. Treatment with PAA was carried 2 h prior to lytic induction and continued during lytic reactivation. Infected iSLK cells that were either left untreated or induced to undergo lytic reactivation with no PAA treatment were used as control. mCherry-ORF45 was used to track cells undergoing lytic reactivation while ORF65, detected with anti-ORF65 antibody followed by anti-mouse Alexa Fluor 488, was used to distinguish late viral lytic stage and effectiveness of PAA treatment. Cells were then stained with anti-UBF (A), or anti-Fibrillarin (B) followed by anti-rabbit Alexa Fluor 647-conjugated secondary antibody. Right-hand columns show the merged output of the nucleolar proteins immunofluorescence with differential interference contrast (DIC) microscopy images.