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. 2015 Sep 2;16(9):1177–1191. doi: 10.15252/embr.201540467

Figure 1.

Figure 1

Oct4 binding to the Sox/Oct motif and specificity for the PORE

  • A, B EMSA (DNA detection) and FP-EMSA (protein detection) of GFP–Oct4 interaction with the Nanog Sox/Oct motif or with PORE motifs, in the absence (A) or presence (B) of mCherry-Sox2.
  • C, D FP-EMSA of binding of Oct4 and Sox2 to the Nanog Sox/Oct motif (C) or to a Sox/Oct motif with a 3-bp insertion (D) between the Oct and Sox components of the Sox/Oct motif. In the presence of Oct4 and Sox2, the bands shift completely from the positions of monomers to the positions of heterodimers in (C), indicating synergistic binding, but not in (D).

Data information: Control: a non-binding GFP-TF fusion protein, O4D: GFP-Oct4 homodimer complex, O4SD: GFP-Oct4/GFP-Sox2 dimer complex on DNA, S2M: GFP-Sox2 monomer on DNA motif, O4M: GFP-Oct4 monomer on DNA motif, FP: free protein, NS: non-specific binding, and FD: free Cy5-tagged DNA motif. All oligonucleotide sequences are listed in Appendix Table S1. n = 3 for (A, B), 2 for (C, D). Source data are available online for this figure.