Figure 4. Ascl2 binding to the human CDX2 promoter.

Chromatin isolated from shRNA-Ascl2/LS174T and shRNA-Ctr/LS174T cells was subjected to immunoprecipitation using H₂O (negative control), IgG antibody (negative control), anti-histone H3 antibody (mAb) (positive control) and mouse monoclonal IgG against Ascl2 (Millipore, MAB4418). The input represents 10% of the DNA used in the immunoprecipitation. A. Five sites in the CDX2 promoter with different numbers of E-box elements (ChIPs 1-5) were tested. B. The final DNA extracts were PCR-amplified using primers. C. The enrichment of the indicated genomic DNA fragments (ChIPs 1-5), the intergenic control (PC) or unspecific binding (blank and NC) was determined relative to the diluted input in three independent experiments.