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. 2020 Mar 6;11:1222. doi: 10.1038/s41467-020-15006-4

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a xDNMT1 pull-downs from native chromatin extracts were analyzed by LC-MS/MS. The volcano plot summarizes the quantitative results and highlights the interacting proteins enriched upon addition of ubiquitin to UbVS-treated extracts. b Xenopus interphase egg extracts were added with sperm chromatin and incubated in the absence or presence of His₆-ubiquitin (58 μM final). Chromatin-bound proteins were isolated and analyzed by immunoblotting using the indicated antibodies. For PAF15 levels in the extracts, see Supplementary Fig. 1f. c Interphase egg extracts were added with sperm chromatin and incubated in the presence of 15 μM aphidicolin (Aph) or in its absence (DMSO). Chromatin-bound proteins were isolated and analyzed by immunoblotting using the indicated antibodies. d PAF15-deleted extracts were supplemented with wild-type xPAF15-Flag₃ and its variants (K18R, K27R, and K18R/K27R). After the addition of sperm chromatin, chromatin-bound proteins were isolated and analyzed by immunoblotting using the indicated antibodies. The extracts were also analyzed by immunoblotting. e PAF15-depleted extracts were supplemented with wild-type xPAF15-Flag₃, its PIP-box mutant (FF/AA), or K18R/K27R mutant (KRKR). After the addition of sperm chromatin, chromatin-bound proteins were isolated and analyzed by immunoblotting using the indicated antibodies. The extracts were also analyzed by immunoblotting. Source data are provided as a Source Data file.