FIG 7.

Overexpression of ORF50 RTA in KSHV-infected Vero cells induces cytoplasmic LANA and ORF59 expression. Vero cells were infected with KSHV for 24 h and then were either mock infected (A and B) or superinfected with BacK50 recombinant baculovirus expressing the KSHV ORF50 RTA (6 h) to activate KSHV replication (C and D); the cells were then cultured for an additional 24 h. The cells were fixed and sequentially stained for ORF59, LANA, and cell nuclei, as described in Fig. 4. Panels A and C show the TO-PRO-3 (blue) and ORF59 (red) fluorescence; panels B and D show the TO-PRO-3 (blue) and LANA (green) fluorescence, with the ORF59-positive nuclei rendered as red spheres. The arrowheads indicate cells with cytoplasmic LANA fluorescence and no ORF59 fluorescence. (E) Cell populations expressing cytoplasmic LANA and nuclear ORF59 were quantitated (number of cells analyzed = 375 [KSHV] and 339 [KSHV+BacK50]). Scale bar, 50 μm.