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. 2014 Feb 18;9(2):e89399. doi: 10.1371/journal.pone.0089399

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© 2014 Makrantoni et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Wild type SLI15 (VMY316), sli15-20A (VMY318) and sli15-20D (VMY320) cells containing CEN5-(tetO)₃₃₆, tetR-GFP, Venus-TUB1 and pMET3-CDC20 were arrested in G1 with α-factor at 26°C and then released to a metaphase block in rich medium (containing 2 mM methionine to deplete Cdc20) for 2.5 h. (A) Representative stills from time-lapse images of live cells. Bi-oriented chromosomes show dynamic splitting and reassociation of sister CEN5s. Green, CEN5 labeled with tetR-GFP; red, Venus-tubulin. (B) Quantification of chromosome bi-orientation in metaphase-arrested cells from multiple time-lapse fields (n = number of cells scored in each category).