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. 2011 Jul 9;39(19):8355–8365. doi: 10.1093/nar/gkr528

Figure 1.

Figure 1.

Dnmt1, UHRF1 and USP7 interact with one another in vivo. (A) USP7 is identified as a new interaction partner of Dnmt1. Dnmt1 was immunoprecipitated from nuclear extracts (Dignam-extract and MNase-extract) and subjected to SDS–PAGE and Coomassie blue staining. Protein bands [(U) USP7, (P) PCNA, (N) UHRF1] specific for the Dnmt1-IP were identified by MS analysis (ctrl: proteinG sepharose only). Precipitated Dnmt1 (D) and the molecular weight marker (M) are indicated. (B) Immunoprecipitation of endogenous proteins of the MNase treated extract with the indicated antibodies. Co-precipitated proteins were detected by Western Blot. 2% of the input (In), 10% of the flowthrough (Ft), 10% of the control beads (PG: ProteinG Sepharose) and 10% of the antibody coupled beads (B) were loaded. (C and D) HeLa S3 nuclear extracts [Dignam-extract (C) and MNase-extract (D)] were applied on a supererose6 gelfiltration column (GE Healthcare). Every second fraction was analyzed on SDS–PAGE following western blot analysis with the indicated antibodies. Load (L), void and the migration of the molecular weight reference proteins are indicated.