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. 2020 Apr 1;219(6):e201907082. doi: 10.1083/jcb.201907082

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© 2020 Jackman et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure S5. — MAD2 recruitment to kinetochores is delayed when MAD1 cannot bind cyclin B1. Related to Fig. 5. (A) Schematic showing how RFP670 was tagged at the N terminus of MIS12 (RHA and LHA refer to right and left homology arms, respectively). (B) PCR of genomic DNA from wild-type RPE1 cells (control), parental RPE cyclin B1-Venus^+/−: Ruby-MAD2^+/−; RFP670-MIS12+/+ cells (Par.), or MAD1 E53/E56K: RFP670-MIS12^+/+ clones 7D2 and 8B12, showing integration of RFP670 into both alleles of MIS12. (C) Maximum projection images of parental RPE cyclin B1-Venus^+/−:Ruby-MAD2^+/−; RFP670-MIS12^+/+cells (Par.) and MAD1 E53/E56K: RFP670-MIS12^+/+ clones 7D2 and 8B12. Left panels show Ruby-MAD2; right panels show RFP670-MIS12. Scale bar, top right panel, 5 µm.