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. 2021 Mar 19;12:1746. doi: 10.1038/s41467-021-21932-8

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© The Author(s) 2021

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. 2 — a Recruitment by the OC complex, starting with the tetramer Mcm5/3/7/4 (lane 2) up to the full heptameric MCM-Cdt1 complex (lane 6). The missing MCM-Cdt1 subunits, in lanes 2–5 and 7 and 8, are labelled with ∆ symbol and displayed as grey hexamers in the cartoon, while black hexamers indicate the subunits present in the assay. Negative controls of the MCM-Cdt1 recruitment are shown in lanes 7 (minus Mcm4) and 8 (minus Mcm3). 20% of the MCM-Cdt1 subunits used in the assay are shown in lane 1. b Two halves of MCM, Mcm5/3/7 and Mcm2/6/4, were isolated from gel filtration in separate runs and they were added separately and tested for recruitment. A constant amount of Mcm5/3/7 was used in combination with an increasing amount of Mcm2/6/4, without or with Cdt1 (lanes 2–4 and 6–8, respectively). The MCM-Cdt1 recruited in lane 8 was also tested for loading (ATP and high salt wash) (L, lane 9). c Recruitment of the MCM-Cdt1 either with Mcm4 wt (lane 1) or with increasing amounts of Mcm4 that lacks its C-WHD (M4∆C, lanes 2–4). d The same as c but with Mcm6 wt and Mcm6 lacking its C-WHD (M6∆C). e Recruitment of MCM-Cdt1 where both M6∆C and M4∆C are included. See also Supplementary Figs. 3 and 4.