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. 2014 Apr 30;4(4):140043. doi: 10.1098/rsob.140043

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© 2014 The Authors. Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/3.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 3. — Eic1 is required for Cnp1^CENP-A assembly, while Eic2 is dispensable. (a) ts mutations in eic1 affect cell viability, while eic2Δ cells show no defects in growth. Five-fold serial dilutions of cells spotted on YES + Phloxine B media and incubated at the indicated temperatures; dead cells stain dark pink. (b) eic1 mutants display reduced Cnp1^CENP-A levels at centromeres. qChIP analyses of Cnp1^CENP-A association with centromeres in the indicated strains when grown at permissive (25°C) versus restrictive temperature (36°C) for 8 h. Enrichment of cc2 DNA relative to the act1 locus is presented. (c) eic1 mutants display sensitivity to TBZ, while eic2Δ cells show no TBZ sensitivity. Five-fold serial dilutions of cells spotted on YES media (untreated) or YES media supplemented with 12.5 μg ml⁻¹ TBZ, and incubated at 25°C. (d) eic2Δ cells display no loss of Cnp1^CENP-A at centromeres. qChIP analyses of Cnp1^CENP-A association with centromeres in the indicated strains when grown at 32°C. Enrichment of cc2 or cc1/3 DNA relative to the act1 locus is presented. Error bars in (b,d) represent standard deviation between at least three biological replicates.