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. 2008 Mar 24;180(6):1115–1131. doi: 10.1083/jcb.200708170

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Copyright © 2008, The Rockefeller University Press

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Figure 4. — Nuc1 is required for localizing condensin at rDNAs. (A) Localization of Cut14-GFP was examined in the nuc1-632 mutant cultured at 36°C for 6 h. (B) Localization of Cut14-GFP was examined in the nuc1-632 mutant with the nucleolar protein Gar2-mCherry. (C) A ChIP assay was performed using a block-release nda3-311 mutant that expressed Cut14-Flag. Nine DNA probes derived from the rDNA repeat unit were used: NTS (N1–N4), ARS, external transcribed sequence, and 18S, 5.8S, and 28S coding sequences. WCE, whole cell extract; IP, immunoprecipitate. The negative control (−) is the strain that did not carry Cut14-Flag. Quantitative enrichment is shown on the right. (D) A ChIP assay was performed using extracts of asynchronous wild-type and nuc1-632 mutant that expressed Cut14-Flag. The wild-type and nuc1-632 mutant were cultured at 26°C and shifted to 36°C for 6 h, and immunoprecipitation was performed. Real-time PCR was used to amplify and quantify DNAs with the PCR primers (N1, N3, lys1 ⁺, and cnt1). Error bars represent SD. Bars, 2 μm.