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. Author manuscript; available in PMC: 2013 Jun 21.
Published in final edited form as: Cell. 2012 Dec 21;151(7):1417–1430. doi: 10.1016/j.cell.2012.11.022

Figure 4. MeCP2 is the major protein that binds 5hmC.

Figure 4

(A) Silver stained SDS-PAGE gel of nuclear proteins from frozen rat cerebella that bound beads coated with unmodified C, 5mC or 5hmC DNA. Arrow pointing at the band that was excised and identified by MS from a Coomassie stained replica gel.

(B) Southwestern blot of the nuclear proteins from cerebella of WT or KO mice (IN) that bound to C or 5hmC- coated beads probed with radioactive 5mC or 5hmC DNA.

(C) EMSA of C, 5mC (M) or 5hmC (H) probes with increasing concentrations (0 to 2 pmol) of recombinant human MeCP2. The arrow points at the MeCP2-dependent low-mobility complexes.

(D) EMSA of C, 5mC (M) and 5hmC (H) probes in presence of 1 pmol of MeCP2 (aa 1-205), 1.2 pmol of MBD1, 0.5 pmol of MBD2, 50 pmol of MBD3 or 2.5 pmol of MBD4. The probes were either reacted with βGT prior to labeling or not (βGT−). Arrows point at protein-dependent DNA complexes.