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. 2007 Dec 13;27(1):111–121. doi: 10.1038/sj.emboj.7601955

Figure 4.

Figure 4

Rad21 remains chromatin bound when cohesin loading is inactivated in S-phase. Cells bearing rad21-HA and the thermosensitive mutations mis4–367 or ssl3–29 were arrested in early S-phase by hydroxyurea (HU) treatment at 25°C and then shifted to 37°C for 2 h in the presence of HU. (A) Rad21-HA association with chromatin was monitored on chromosome spreads by immunofluorescence before and after the shift to 37°C. Scale bar, 5 μm. (B) Rad21-HA fluorescence was measured in 50–100 nuclei. The error bars show the confidence interval of the mean with α=0.05. (C) DNA content analysis and septation index (SI) show that cells remained arrested with close to 1C DNA content throughout the experiment.