FIG 5.

Immunofluorescence assay (IFA) to determine KSHV protein expression and FISH to examine viral DNA replication. (A and B) Cells of the SLK-iBAC-gfpK52 cell line were transfected with a SIRT6-expressing plasmid; 24 h later, the cells were treated with 100 nM doxycycline. Twenty-four hours later, the cells were fixed for IFA to detect the KSHV protein RTA. The pictures were taken under a confocal microscope at magnifications of ×10 (A) and ×40 (B). (C) FISH was performed to examine DNA replication. (D) The proportions of RTA-positive cells counted among total cells, SIRT6-positive cells, and SIRT6-negative cells were determined. (E) The proportions of DNA replication-positive cells among total cells, SIRT6-positive cells, and SIRT6-negative cells were determined. A Student's t test was performed to calculate statistical significance. **, P < 0.001. DAPI, 4′,6-diamidino-2-phenylindole.