(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.