1. Cdx2 and Oct4 bind directly to Xist intron 1 and Xite in vitro.
Cdx2 and Oct4 bind directly to Xist intron 1 and Xite in vitro. (A) Gel shift of Xist intron 1 (Int1), using recombinant Cdx2 (rCdx2). Arrow, protein–DNA shift. Comp, cold competitor added at 100-fold molar excess; Mut, mutation. (B) Gel shift of the Xite motif using rCdx2. (C) Gel shift of Xist intron 1 and Xite, using recombinant Oct4 (rOct4). (D) Gel shift of Xist intron 1 motif using rOct4 and rCdx2. Two times (2×) and three times (3×),molar excess of rOct4 are shown. mCdx, mutated Cdx2-binding site; mOct-mCdx, mutated Cdx2-Oct4–binding sites. (E) qRT-PCR for Xist RNA, with levels normalized to β-actin levels. Mean ± SD is shown. ES, Cdx2-transgenic ES clones without TX induction of Cdx2. iTS, Cdx2-transgenic clones transdifferentiated to TS cells by TX induction of Cdx2 for 1 (d1) or 2 (d2) days. (F) Allele-specific gChIP analysis for Cdx2 and Oct4 binding from indicated samples.
Jennifer A. Erwin et al. Genetics 2012;192:
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