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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 8. — Proper segregation of sister kinetochores and rDNAs requires condensin. (A) Cen2-GFP was used to examine the segregation phenotype of centromeric DNA in the condensin mutant cut14-208. The wild type, mis6-302, and mis12-537 were used as the control strains. These were first grown at 26°C and were shifted to 36°C for 2 h (wild type and cut14-208) or 8 h (mis6-302 and mis12-537). The frequency of equal or unequal segregation is shown at the bottom of the images. (B) Kymographs for Cen2-GFP are shown for the wild type and cut14-208, which also expressed Sad1-RFP for the SPB marker. Images were taken at 10-s intervals. The frequency of each phenotype is indicated in parentheses. The arrow in the bottom panel (cell 2) shows the two closely situated splits of Cen2-GFP signals upon the presumed onset of anaphase. (C) The rDNA repeat unit was used for the FISH probe in the wild-type and condensin mutant cells. DAPI and anti-Sad1 antibodies were used to stain DNA and to immunostain the SPBs, respectively. (D) Steps required for condensin to be accumulated at mitotic kinetochores and rDNAs in S. pombe. Cut15, importin α; Cut17, Bir1/survivin; Ark1, aurora; Cnp1, CENP-A; Mis6, CENP-I; Mis13, hMis13; Nuc1, pol I largest subunit; Acr1, accumulation of condensin at rDNA. Bars: (A) 5 μm; (B), 2 μm; (C) 10 μm.