Xenbase Image ID: 157220

Figure 2. Usp7 forms a stable complex with Dnmt1 in Xenopus egg extracts. (A) Dnmt1 was immunoprecipitated from interphase HSS using specific antibodies. The associated polypeptides were identified by mass spectrometry. The resultant immunoprecipitates were subjected to SDS-PAGE and stained with a SyproRuby; the peptides corresponding to Dnmt1 and Usp7 are indicated. The numbers of unique spectra identified for each protein are shown in the table. (B) Immunoprecipitates using anti-Dnmt1 (lane 3), anti-Usp7 (lane 4), anti-Uhrf1 (lane 5) antibodies, or control IgG (lane 2) as well as egg extracts (lane 1) were subjected to immunoblotting using the antibodies indicated. (C) Sequence alignment of the KG linker of Dnmt1 across different species. Red denotes residues mutated in a 4KA mutant used in this study. (D) FLAG-tagged wild-type or a 4KA mutant of recombinant Xenopus Dnmt1 was introduced into Xenopus egg extracts. Recombinant or endogenous Usp7 was immunoprecipitated using either anti-FLAG-M2 agarose beads or protein A agarose beads conjugated with anti-Usp7 antibodies, respectively. Immunoprecipitates were then subjected to immunoblotting using the antibodies indicated. Source data are available online for this figure. Image published in: Yamaguchi L et al. (2017) © The Author(s) 2017. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

Return to previous page