FIGURE 6.

LANA Arg-20 mutations affects KSHV chromatin binding during latency. A, KSHV wt-LANA whole genome tiling array is shown. LANA ChIP DNAs were prepared from K-Rta-inducible BCBL-1 cells during latency. The immunoprecipitated DNA was probed against a KSHV tiling array as described under “Experimental Procedures.” The graph depicts the ratio of LANA/input signal across the KSHV genome. The entire KSHV genome is annotated at the bottom of the figure. The arrows denote ORFs evaluated by ChIP and/or RT-qPCR in the 293T BAC stable cell cultures. Several regions of the KSHV genome are indicated above the graph for reference. Ba, validation of the anti-LANA antibody in LANA ChIP assay is shown. ChIP DNA samples were prepared as described under “Experimental Procedures” and immunoprecipitated with anti-LANA antibody or normal rat IgG. DNA associated with each immunoprecipitate was analyzed by qPCR using primer pairs against the ORF73 promoter. Values are reported as the ratio of the Ct values of the amplified DNA immunoprecipitated relative to the amount of input DNA amplified. b, ChIP analysis shown is. ChIP DNA was prepared from each BAC stable cell line during latency. ChIP DNA associated with each immunoprecipitate was analyzed by qPCR using primer pairs directed against the ORFs listed. Values are reported as the ratio of the Ct values of the amplified DNA immunoprecipitated with anti-LANA antibody relative to the amount of input DNA amplified.