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. 2019 Sep 4;10:3987. doi: 10.1038/s41467-019-11773-x

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© The Author(s) 2019

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. 9 — SENP6 knockdown leads to the depletion of multiple CCAN proteins from chromatin. a U2OS cells were transfected either with a pool of four siRNAs against SENP6 (siSENP6) or a pool of four nontargeting siRNAs (NTP). Chromatin fractions were isolated and proteins were analyzed by mass spectrometry. Volcano plot showing all identified proteins within the NTP treated sample compared with the siSENP6 treated sample. All identified inner kinetochore proteins are represented by red circles. n = 4 independent experiments. b Samples from panel a were analyzed by immunoblotting using antibodies against several inner kinetochore proteins, as well as β-tubulin and histone H4 as control for efficient fractionation. Source data are provided as a Source Data file. c Model of SENP6 regulation of CCAN proteins. SENP6 deSUMOylated CCAN proteins as well as Mis18BP1. SENP6 depletion leads to RNF4-mediated degradation of Mis18BP1 and consequently the failure of CENP-A to accumulate at the centromere. Reduced CENP-A levels as well as accumulated SUMO chains on other CCAN family members hinder efficient CCAN protein complex formation at the centromere