Association of STAT3 with SIE/GAS sites identified inDNMT1gene promoter 1 and enhancer 1 in vitro. (A) Map of the DNMT1 gene promoter and enhancer region with highlighted STAT3-binding sites and corresponding DNA oligonucleotide probes used in EMSA and primer sets used in ChIP (for comparison, see Figure 4). (B) EMSA with DNA oligonucleotides that corresponded to promoter 1 (P1) and enhancer 1 (E1) STAT3-binding sites with nuclear protein extracts from malignant T cells 2A and normal PBMCs as control. (C) Binding of the 2A T-cell line protein extracts (protein) to the enhancer 1 and promoter 1 STAT3-binding sites in the presence and absence of the unlabeled (cold and SHP-1) probes. (D) Binding of the 2A and 2B T-cell line protein extracts to promoter 1 and enhancer 1 STAT-binding sites either wild-type (P1 and E1, respectively) or with mutated (M) TT and AA pairs by substitution with the CC pairs. (E) Supershift EMSA with the anti-STAT3 and control anti–anaplastic lymphoma kinase (ALK) antibody.