Promoter analysis and expression of Id2 and Tcf4 in DCs. (A) Schematic diagrams showing the location of putative STATx (open box) and C/EBPβ (gray box) consensus sites in the murine and human Id2 (left) and Tcf4 (right) promoter regions. Arrows indicate the approximate locations of primers used for STAT ChIPs. Primer sequences are listed in supplemental Table 1. (B) FACS-purified pDC and cDC subsets were analyzed for Tcf4 and Id2 mRNA expression by quantitative PCR, as indicated. DC purification strategies are shown in supplemental Figure 1. Results for Tcf4 and Id2 were normalized to Rpl13a. Results are shown as mean ± SEM of 3 independent experiments. (C-D) Liver CD103⁺ DCs and BM pDCs were purified from mice 7 days after GM-CSF or Flt3L HGT by FACS. Histone H3 modifications at the Id2 and Tcf4 proximal promoters were analyzed by ChIPs using antibodies against H3K9ac, H3K4me3, or H3K27me3; results were normalized to data from ChIPs with total H3 antibody. Normalized data were plotted versus the corresponding predicted ChIP amplification products from the Id2 and Tcf4 2 kb proximal promoter regions, as indicated by the schematic diagrams in the bottom panel, which include the approximate location of putative transcription factor binding sites. Results from CD103⁺ DCs (○) or pDCs (●) are presented as mean ± SEM of 3 independent experiments for individual modifications, as indicated.