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

Figure 5.

Figure 5

Pds5’s function in cohesion maintenance remains essential in the absence of both Eso1 and Wpl1. (A) Kinetics of Rad21 dissociation from G2 chromosomes upon inactivation of the cohesin loader. Cycling cells were shifted to 37 °C to shut off cohesin loading (mis4-367) while keeping cells in G2 (cdc25-22). Chromatin-bound Rad21 was measured by nuclear spreads (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 showing that cells were predominantly in G2 before the temperature shift and remained in G2 throughout the experiment. (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.