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. 2017 Jun 15;28(12):1701–1711. doi: 10.1091/mbc.E16-12-0846

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© 2017 Lawrimore et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — Pericentric cohesin responds to DNA damage. (A) Pericentric cohesin in metaphase with Smc3-GFP and Spc29-RFP in untreated, phleomycin, Zeocin, and a sDSB induced by HO endonuclease from the GAL10 promoter, target site at the MAT locus (KBY6050). Bar, 1 μm. (B) Cohesin sagittal width (schematic below A; red, spindle poles; green, cohesin; black, spindle microtubules) under DNA damage. Untreated (n = 24), plus phleomycin (n = 70), GALHO no induction (n = 38), and GALHO induction (n = 58) caused significant expansion (asterisks) of cohesin (Student’s t test, p < 0.05) compared with untreated (n = 50) and no GAL-HO (n = 20). (C) Cohesin barrel fluorescence intensity over arm untreated (n = 50), plus phleomycin (n = 40), plus Zeocin (n = 43), GALHO (n = 40), and I-SceI (n = 29). Student’s t test showed a difference between untreated and damaged cells (p < 0.05). (D) Metaphase cells showing three different kinetochore protein (Ame1-GFP) phenotypes: two spots (blue), one elongated (purple), and more than two spots (red). Asterisk denotes LacO/LacI in KBY 6047. Bar, 1 μm. (E) Bar graph showing the distribution of Ame1-GFP phenotypes in metaphase for WT cells (n = 92) plus phleomycin (n = 101), plus Zeocin (n = 153), and plus I-SceI (n = 123). Error bars are SEM.