FIG 8.

Chromatin immunoprecipitation (ChIP) assay to determine the interaction of SIRT6 with KSHV DNA. (A) The anti-SIRT6 antibody was used to detect SIRT6 in SLK-iBAC-gfpK52 cells with or without doxycycline for 48 h. The whole-cell lysates were applied to the Western blot assay to examine the indicated proteins. (B) Cells of the SLK-iBAC-gfpK52 cell line were treated with or without doxycycline for 48 h, fixed with 4% formaldehyde, sonicated to shear the DNA to fragments over a range of 200 to 400 bp in size, and aliquoted into three groups: input, normal mouse IgG incubation, and incubation with anti-SIRT6. The pulled down DNA was detected by PCR using primers designed for the ori-Lyt promoter, the ORF50 promoter, or ORF50 gene-encoding sequences (ORF50 gene). The DNA amount was compared to the input amount, and the result is shown as a percentage.