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. 2016 Jun 7;6(8):2655–2663. doi: 10.1534/g3.116.031674

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Copyright © 2016 Shwartz et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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RID mutations do not affect loader integrity. (A) Haploid YMS1003 (SCC2-12Myc), YMS1016 (SCC2-12Myc SCC4-3V5), YMS1017 (SCC2-12Myc scc4-L305ins-3V5), YMS1018 (SCC2-12Myc scc4V485ins-3V5), YMS1019 (SCC2-12Myc scc4L490ins-3V5), and YMS1020 (SCC2-12Myc scc4S505ins-3V5) were grown to midlog phase in YPD medium at 23°. Cells were lysed, and the protein extracts were subjected to IP against the V5 tag (Scc4). Precipitated Scc2 was analyzed by Western blot using antibodies against Myc. (B) Strains YIO002 (scc4-4), YMS1004 (SCC4-3V5 scc4-4), YMS1006 (scc4L305ins-3V5 scc4-4), YMS1007 (scc4V485ins-3V5 scc4-4), YMS1005 (scc4L490ins-3V5 scc4-4), and YMS1008 (scc4S505ins-3V5 scc4-4) were processed for ChIP analysis. V5 tagged Scc4 proteins were immunoprecipitated (n = 3). Precipitated DNA was analyzed by quantitative PCR. Analysis of centromere III is shown. (C) Analysis of centromere IV is shown.