Supplemental Figure 1 Kang et al. Probe Octamer MORE NE - WT S186D - WT S186D MMS - - + - + - - + - + 2 NS 1 P 1 2 3 4 5 6 7 8 9 10 Figure S1. Effect of Oct4 S186 aspartic acid mutation on stress induced MORE dimerization. EMSA using nuclear extracts from Oct1 deficient MEFs infected with Oct4 WT or S186D retroviruses. Fibroblasts were treated with 200ug/ml MMS for 4hr and collected immediately. P: probe. NS: nonspecific. 1: monomeric Oct4 complex. 2: Oct4 dimer. Supplemental Figure 2 Kang et al. A B Probe Octamer TMFORE NE - + + - + + IR - - + - - + 2 1 P 1 2 3 4 5 6 Supplemental Figure 2 (cont.) Kang et al. C Probe Oct MR 2XMR TMFORE NE + + + - + + IR - - - - - + 4 2 1 1 2 3 4 5 6 Supplemental Figure 2 (cont.) Kang et al. D -Oct4 -Oct4 - α Foxo4 - α Piwil2 IP IP INPUT INPUT E Foxo4 Piwil2 (Mili) Taf12 Figure S2. Identification of a new Oct protein binding element. A) Screenshot from the UC Santa Cruz Genome Browser (http://genome.ucsc.edu) of the human Taf12 gene regulatory region on chromosome 1. B) Oct4 dimerization on a TMFORE sequence derived from the Taf12 regulatory region following IR treatment. P: probe. 1: monomeric Oct4. 2: dimeric Oct4. C) EMSA using HeLa nuclear extracts and simple octamer (Oct), MORE (MR), 2XMR or Taf12 probes of equal size. Arrow at right shows monomeric, dimeric, and tetrameric probe occupancy. The free probe was run off the gel to resolve bound species. D) Alignment of three mouse regions is shown. All sequences were highly conserved to human. Yellow box: conserved sequences between all three regions, Blue box: conserved sequences between two regions. Arrows indicate positions of octamer half-sites. Yellow arrows: POU-homeodomain; blue arrow: POU- specific domain. E) ChIP assay using mouse ES cells and Oct4 antibodies together with Foxo4 and Piwil2 (Mili) genomic promoters. Supplemental Figure 3 Kang et al. A B Chr11:56,849,044-56,849,088: TNKS1BP1 Chr20:35,589,940-35,589,984: BLCAP C D ChrX:149,902,323-149,902,367: HMGB3 Chr11:14,337,314-14,337,358: RRAS2 Supplemental Figure 3 (cont.) Kang et al. E F chr12:120,389,591-120,389,635: FBXL10 chr9:34,448,591-34,448,635: c9orf25 G H chr19:54,835,235-54,835,279: RRAS Chr19:18,494,095-18,494,139: ELL Supplemental Figure 3 (cont.) Kang et al. I Chr7:44,754,922-44,754,966: ZMIZ2 Figure S3. Characterization of new Oct1 targets regulated by MORE identified by ChIPseq. A-I) Screen shots from the UC Santa Cruz Genome Browser of MORE regulated regions identified by ChIPseq. Up per panel: red arrowhead indicates the position of MORE. Lower panel: yellow boxes contain highly co nserved MORE sequences..