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Fig. 3. Localization of macroH2A in wild-type and demethylated ES cells. (A) Representative ES cells subjected to macroH2A immunofluorescence (green) and DAPI staining of DNA (blue). J1 is a wild-type ES cell line. N/N, S/S and C/C denote ES cells with Dnmt^N/N, Dnmt^S/S and Dnmt^C/C genotypes, respectively. ES cell line B9 was derived by introduction of a Dnmt1 BAC into cell line into Dnmt1^C/C cells resulting in partial remethylation of genomic CpG dinucleotides. (B) The same ES cell lines immunostained with CREST autoimmune serum, which recognizes centromeric kinetochore proteins including the histone H3 variant CENP-A. (C) Representative immunostainings of mitotic J1 and C/C ES cells using ${alpha}$ -macroH2A and CREST serum. Note that in J1, macroH2A and kinetochore immunostaining is distinct, while in C/C cells, immunostaining is nearly coincident.

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