Figure 7.

Chromatin-immunoprecipitation (ChIP) analysis of HEK 293T cells demonstrates that specific sequences near TSS II bind IRF-3. HEK 293T cells were transfected with pEGFP-C1 (hIRF-3) to overexpress IRF-3 or pEGFP-C1 control vector. After 48 hr cells were fixed as described in Methods and processed for ChIP analysis. Antibody to IRF-3 or rabbit (Rb) IgG was used to precipitate chromatin-bound IRF-3. A. DNA sequences amplified near TSS II and downstream (primers indicated above bars and given in Table 1). Negative numbers indicate base pairs upstream of the start codon. B. Quantities of precipitated DNA from cells transfected with pEGFP-C1(hIRF3) or pEGFP-C1 were determined by RT-PCR. The DNA precipitated with anti-IRF-3 antibody was compared to similar DNA precipitated with Rb IgG, which was considered baseline. Primers for each amplified sequence are given below.