IRF5 is recruited to the 5′ upstream and 3′ downstream region of TNF. (A) Schematic of the TNF locus. Protein coding and noncoding exons are shown in black and white. Putative ISREs are allocated as white ovals; κB sites are shown as black circles. The approximate amplicon size of primer sets spanning the TNF locus (A to L) are indicated by black lines. CO indicates a control primer set containing neither an ISRE nor a κB site. (B-C) HEK-293–TLR4-Md2/CD14 cells were left unstimulated or stimulated with LPS (1 μg/mL) for 4 hours and 24 hours and analyzed by ChIP with antibodies specific to IRF5 (B) or RelA (C). (D-E) MDDCs were left unstimulated or stimulated with LPS for 1 hour and 4 hours, followed by ChIP with antibodies specific to IRF5 (D) or RelA (E). (F) Corecruitment of RelA and IRF5 to region H was assessed by re-ChIP analysis with antibodies against RelA followed by IRF5-specific antibodies. (B-F) Data show mean percentage input relative to genomic DNA (gDNA) plus or minus SD of a representative experiment. −AB indicates a no-antibody control.