Figure 7: The PLZF-HDAC3-NF-κB p50 complex is a transcriptional repressor.

(a–c) Measure of transcriptional factor binding to gene promoters by detecting PCR amplicons from the IL-12p40 and TNF-α promoters after (a) ChIP of products with anti-PLZF, -NF-κB p65, -p50, -HDAC3, -p300 or, as a control, an IgG antibody, from WT and Zbtb16^−/− BMMs treated with LPS (10 ng ml⁻¹) for the times indicated. (b) Re-ChIP of products initially IP from WT and Zbtb16^−/− BMMs treated with LPS (10 ng ml⁻¹) for the times indicated. The initial IP used anti-HDAC3 or -NF-κB p50 antibodies that were then Re-ChIP with anti-NF-κB p50, -p65 or -HDAC3, as indicated. The amplicons from the total lysates for each promoter are shown at the bottom (Input). (c) Graphs quantitating sequence tags from the Tnf-α promoter enriched by ChIP with anti-HDAC3, -NF-κB p50, -p65 and -p300 antibodies from WT and Zbtb16^−/− BMMs treated with LPS (10 ng ml⁻¹) for the times indicated. The graphs show the mean±s.e.m. (*P<0.05 by Student’s t-test, n=3).