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. Author manuscript; available in PMC: 2012 Dec 23.
Published in final edited form as: Cell. 2011 Dec 23;147(7):1498–1510. doi: 10.1016/j.cell.2011.11.054

Figure 6. Mbd3 directly binds to hydroxymethylated DNA in vitro.

Figure 6

(A) Silver stain showing tandem affinity purification from untagged or Mbd3-6His-3XFLAG ES cells.

(B) Western Blot of purifications described in (A) for indicated proteins. Mta1 is a component of the Mbd3/NURD complex. Preliminary mass spec results also identified most other known NURD subunits in this purification (not shown).

(C) EMSA assay using Mbd3/NURD complex and DNA probes containing unmodified cytosine (C), methylated cytosine (5mC) or hydroxymethylated cytosine (5hmC).

(D-E) EMSA assay using recombinant mouse Mbd3 (D) or recombinant Mbd1 methyl-binding domain (E), and various modified probes as in (C).

See also Figure S6.