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. 2015 Jun 30;43(20):e132. doi: 10.1093/nar/gkv670

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

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

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Figure 9. — Pericentric cohesin viewed by calibrated ChIP-seq and cell images. (A) Cohesin's peri-centric distribution in G2/M phase cells. The number of reads per base pair (Y-axis) in the 30 kb regions either side of the CDEIII element for each chromosome (X-axis) were averaged and calibrated to reveal the average cohesin distribution around the centromere. The data was extracted from ChIP-seq of cells at 60 min. after released from G1-arrest in Figure 5. The ‘5 kb’ peaks are marked by red boxes and centromere-proximal cohesin marked by blue box. (B) High-resolution image of cohesin in G2/M phase. Tetraploid cells containing 64 chromosomes with endogenous Scc1 tagged with EGFP (K18719) were exponentially grown in YPD at 25°C and fixed with 2% of formaldehyde. The cells were observed under high-resolution OMX microscope. The lateral view is shown on the top panel and the transverse view on the bottom one. (C) Bi-orientation of sister kinetochores and peri-centric cohesion mediated by ‘5 kb’ peaks (red boxes) in G2/M phase cells. Sixty-four such bi-oriented chromosomes will be clustered around pole to pole microtubules, generating the cohesin barrels shown in (B). Chromosome axes – the black fibre. Kinetochores – mauve balls. Centromere-proximal cohesin - blue box. Pole to pole microtubules – green cylinders.