Figure 7: Binding of Klf5 to Dmp1 promoter.

(a) Scheme represents of 2 potential Klf5 binding sites in human Dmp1 promoter. (b) Conserved non-coding sequences of Dmp1 gene promoters in three mammalian species, mouse, rat and human are retrieved using Vista plots for comparative genomic analysis of gene promoters. The pink regions represent peaks of conserved nucleotide sequences with a minimum of 70 homologies. Many of the conserved non-coding sequences are in these regions. The scale on the y-axis goes to from 50 to 100% homology. Two ³²P-labeled double-stranded Dmp1 probes (Table 1) were generated for EMSAs. These included Dmp1-site 1 and Dmp1-site 2. EMSAs were carried out using nuclear extracts obtained from iMDP-3 cells. (c, lane 1) free Dmp1-site 1 probes only; (c, lane 2) the binding of nuclear extracts to Dmp1-site 1 probe; (c, lane 3) the probe with nuclear extracts and 100× cold Dmp1-site 1 oligo; (c, lane 4) the probe with nuclear extracts and 100× cold Klf consensus sequence oligo; (c, lane 5) the probe with nuclear extracts and 1 μg Klf5 antibody; (c, lane 6) the probe with nuclear extracts and 2 μg Klf5 antibody; (c, lane 7) free Klf consensus sequence probe only; (c, lane 8) the binding of nuclear extracts to the Klf probe; (c, lane 9) the probe with nuclear extracts and 100× cold Klf consensus sequence oligo; (c, lane 10) the probe with nuclear extracts and 100× cold Dmp1-site 1 oligo; (c, lane 11) the probe with nuclear extracts and 1 μg Klf5 antibody; (c, lane 12) the probe with nuclear extracts and 2 μg Klf5 antibody. (d, lane 1–12) Dmp1-site 2 and Klf probes.