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. 2012 May 29;13(7):645–652. doi: 10.1038/embor.2012.72

Figure 4.

Figure 4

The absence of a stable cohesin fraction in pds5Δ is not rescued by wpl1 deletion. Cycling cells were shifted to 37 °C to shut off cohesin loading (mis4-367) while keeping cells in G2 (cdc25-22). (A) Kinetics of Rad21 dissociation from chromatin by nuclear spreads. Fluorescence intensity was measured for 50–100 nuclei per sample. Error bar=95% confidence interval of the mean with α=0.05. (B) Cell survival over time. Error bar=s.d. from three experiments. (C) Cell cycle staging before and after the temperature shift. (D) ChiP assay showing the amount of Rad21-PK bound at the indicated loci, shortly (15 min) and 3 h after the 37 °C temperature shift. Error bar=s.d. from four ChiPs. (E) Cytological analysis of cells undergoing their first mitosis upon release from the cdc25-22 arrest. Green: tubulin, red: DAPI. Bar=5 μm. The frequency of abnormal anaphases was determined as in Fig 2 from the examination of 80–100 anaphases per sample. ChIP, chromatin immunoprecipitation; DAPI, 4,6-diamidino-2-phenylindole.