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. 2011 Feb 2;6(2):e16627. doi: 10.1371/journal.pone.0016627

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Frauer et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A–B) Subcellular localization (A) and binding kinetics (B) of GFP-CXXC^Dnmt1, GFP-CXXC^Dnmt1KF/AA, GFP-CXXC^Tet1 and GFP in mouse C2C12 myoblasts. Localization and binding kinetics were independent from the cell cycle stage (Figures S2 and S5 in File S1). Arrowheads in (A) point to nucleoli. Scale bar: 5 µm. Binding kinetics were analyzed by FRAP. (C) DNA binding specificity of the Dnmt1 and Tet1 CXXC domains. GFP, GFP-CXXC^Dnmt1, GFP-CXXC^Dnmt1KF/AA and GFP-CXXC^Tet1 were pulled down from extracts of transiently transfected HEK293T cells and incubated with fluorescent DNA substrates containing no CpG site or one central un-, hemi- or fully methylated CpG site in direct competition (noCGB, UMB, HMB, FMB, respectively). Shown are the mean DNA/protein ratios and corresponding standard errors from 5 (GFP), 4 (GFP-CXXC^Dnmt1 and GFP-CXXC^Dnmt1KF/AA) and 2 (GFP-CXXC^Tet1) independent experiments. * P = 0.01; ** P = 0.007; ***P = 0.001.