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. 2016 Apr 15;44(14):6676–6692. doi: 10.1093/nar/gkw252

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© The Author(s) 2016. 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 3. — Cohesin at Mis4+/Rad21+ loci is stabilized by Eso1. (A) ChIP-qPCR analysis showing Psm3-GFP enrichment at selected CARs in WT and eso1-H17 cells at 25°C. No tag strain was used to assess the background levels. *P < 0.05, two-tailed, paired Student's t-test for Psm3-GFP compared to background levels. Error bars represent SD, n = 3. (B) ChIP-qPCR analysis showing Psm3-GFP enrichment at selected CARs in WT and eso1-H17 cells at 37°C. No tag strain was used to assess the background levels. *P < 0.05, two-tailed, paired Student's t-test for Psm3-GFP compared to background levels. Error bars represent SD, n = 3. (C) ChIP-qPCR analysis showing Psm3-GFP enrichment at centromeric dg repeat (CAR VI) in WT and eso1-H17 cells at 25 and 37°C. Error bars represent SD, n = 3. (D) Left panel: western blot showing Eso1-GFP protein levels in G2/S phase cells. Whole cell extracts were prepared from cycling cells (G2), cells arrested in early S-phase with Hydroxyurea (HU) and subsequently released into G2 at indicated time points. No tag is a control for anti-GFP antibody specificity and Histone H3 is used as a loading control. Right panel: western blot showing Eso1-GFP protein levels in G1 synchronized cells through to G2 at respective time points. H3 is used as a control. (E) ChIP-qPCR analysis showing Eso1-GFP enrichment at selected CARs in G2 synchronized cells (cdc25-22, 37°C, 4 h). No tag strain was used to assess the background levels. * P < 0.05, two-tailed, paired Student's t-test for Eso1-GFP compared to background levels. Error bars represent SD, n = 3.