A map of transcriptional complexes on DPPA4, a known target of POU5F1 (Oct4). (A) The oligonucleotide pool was subjected to binding assays to identify specific protein DNA complexes by coimmunoprecipitation with SOX2, POU2F1 (Oct1), POU5F1 (Oct4), and NANOG antibodies. Specific oligonucleotide enrichment in the bound fraction was analyzed by two color microarray (bound vs. starting pool). The oligonucleotides were mapped back to genomic coordinates (x-axis), and their average log10 red/green ratio (enrichment) is plotted on the y-axis. UCSC known gene annotation of the DPPA4 indicates transcription and translation start sites and a portion of the first intron of the DPPA4 gene. Dotted vertical lines mark the boundaries of complexes identified in the J1 extracted. (B) The enrichment of DPPA4 and a control region (beta-ACTIN) by chromatin immunoprecipitation with anti-POU2F1 (anti-Oct1), anti-POU5F1 (anti-Oct4), and irrelevant (p53) antibodies measure binding in vivo in J1 ES cells. No antibody and input controls are labeled on the figure. ChIP and sequential ChIP (Re-ChIP) performed on previously identified POU2F1 (Oct1) targets are labeled similarly.
