FIG 7.

Deletion of the CTRL2 insulator did not affect Rad21 enrichment at 28 dpi. ChIP assays using the Rad21 antibody or the IgG control were done on mouse TG infected with the ΔCTRL2 virus at 28 dpi. Subsequent qPCR using primers and custom probes specific for nucleotide regions within 100 to 150 bp of the CTCF insulator sites was performed on both the antibody aliquot and the IgG control. Relative copy numbers in the B, I, or IgG fraction were determined from the equation for the standard curve specific to the primer/probe set and then normalized to IgG. B/I ratios for each site were divided by the IgG/I normalized value so that comparisons between the ΔCTRL2 and wt 17Syn+ virus are graphed as fold enrichment relative to IgG. The bar graph is presented as the average fold enrichment for all 5 biological replicates at 28 dpi for the wt and mutant viruses. One-way ANOVA on independent samples was used to determine statistical significance between fold changes relative to IgG between the two viruses.