Figure 6.

KLF4 bound to mCpGs at enhancer regions to activate LMO7 expression A. Schematic structure of the LMO7 gene locus showing KLF4 binding sites at the 5ʹUTR flanking exon 2 with ~10 kb apart. B. H3K27ac ChIP-seq analysis indicated the enrichment of the enhancer mark at the two KLF4 binding sites of the LMO7 gene, which are also highly methylated according to our whole-genome bisulfite sequencing data analysis (dotted). C. Primers were designed to span the two KLF4 binding sites of the LMO7 gene and to amplify the predicted linker site of the 3D loop. A predicated PCR fragments with the size of 311 bp was enriched in KLF4 WT expressing cells, but not in KLF4 R458A expressing cells. D, E. Sanger sequencing confirmed that the PCR fragment completely matched our putative loop with the linker sequencing as shown in D.