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Fig. 4. Nuclear localization of telomeres in cells expressing byr1DD. h– wild-type (TG309) cells (A) and h– spk1{Delta} (TG335) cells (B) carrying pREP81-byr1DD were cultured in liquid EMM2 for 27 hour at 30°C to induce ectopic meiosis, and observed by fluorescence microscopy without fixation. The SPB (red) was stained with CFP-Sad1, and telomeres (green) were stained with Taz1-GFP. Bright-field images of the same cells are also shown (Phase). Scale bars: 5 µm. (C) Time course of cell shape alteration (`altered shape') and telomere clustering (`tel clustering') after induction of byr1DD in h– wild-type (TG309) cells and h– spk1{Delta} (TG335) cells carrying pREP81-byr1DD. To calculate the percentage of cells with altered shape, at least 130 cells were scored for each time point. To calculate the percentage of cells with telomere clustering, at least 50 cells were scored for each time point. (D) h– wild-type (CRL239) cells carrying pREP81-byr1DD were induced to undergo ectopic meiosis in liquid EMM2 at 30°C, and fixed at 20 hours after induction. The SPB (red) was stained with anti-Sad1 antibodies; telomeres (green) and centromeres (blue) were stained by FISH using cos212 and pRS140 as DNA probes, respectively. The nucleus (white) was stained with DAPI. Scale bar: 5 µm.





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