Figure 5.

Oct4 complex site binding is IR-inducible. (A) Alignment of DNA-binding domain amino acid sequences of human Oct1, Oct2, and Oct4. Homologous amino acids are highlighted. Oct1 S335 and S385 and the homologous Oct2/Oct4 serines are boxed. (B) EMSA using nuclear extracts from Oct1-deficient MEFs infected with retroviruses encoding murine Oct4 or uninfected controls. (NS) Nonspecific; (ND) not determined. Probes: (Oct) Simple octamer; (MR) MORE. (Inset) Oct4 Western blot using input extracts. (C) (Top panels) Screenshots of mouse Bmp4 locus intronic region. The MORE is marked with an arrow. Mammalian conservation is also shown. (Below) Higher-resolution image showing the MORE sequence and conservation. (Bottom panel) ChIP assay using mouse ES cells treated with IR and a primer pair spanning the Bmp4 MORE. (D) Microarray output tracks along the human Taf12 locus. Log₂ microarray intensities are shown for library oligonucleotides. IR-treated ES extracts were prepared at 0-, 1-, and 3-h time points. (E) (Top panels) Oct1 and Oct4 ChIP assays using mouse ES and human HeLa cells and genomic oligonucleotides encompassing the enriched Taf12 region. (Bottom panels) ChIP time course showing Oct4 and Oct1 association with the Taf12 site following IR exposure of mouse ES cells and HeLa cells.