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. Author manuscript; available in PMC: 2016 Jan 20.
Published in final edited form as: J Immunol. 2014 Jul 25;193(5):2574–2586. doi: 10.4049/jimmunol.1400833

Figure 4.

Figure 4

Noncanonical NF-κB subunits RelB and p52 regulate IDO expression via direct binding to the IDO promoter region in MDSCs. (A) Four detective biotinylated probes covering 11 putative sequences were synthesized. (B) Detective probes were incubated with MDSCs nuclear extracts with or without competitive probes to test the specificity of NF-κB-IDO binding. Forty fold dose of competitive probes was used to avoid NF-κB binding to the biotinylated detective probes. * P<0.05. (C1) Binding of p52 and RelB to five regions in the IDO promoter containing 6 putative noncanonical NF-κB binding sequences was measured by ChIP-PCR assay. Regions amplified are highlighted, bracketing putative noncanonical NF-κB binding sequences. MiaPaCa2 cells served as the positive control of p52 and RelB binding in skp2 gene. All products were analyzed on agarose gels. (C2) Quantitative values are calculated using the ratio of ChIP PCR product to Input PCR product (relative ChIP value, enrichment ratio). * P<0.05.