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Fig. 4. Anaphase spindle elongation was slow and not complete in scn1-17 mutant. (A) The spindle index of wild-type and scn1-17 was measured after transfer to 20°C. The short spindle (the metaphase spindle length 
2.5 µm) did not accumulate in scn1-17. (B) Cdc13 (mitotic cyclin in fission yeast) is degraded in aberrant anaphase in scn1. The scn1 mutant carrying chromosomally integrated cdc13-GFP gene was cultured at 20°C for 9 hours in the liquid EMM2, harvested and stained with Hoechst H33342 (DNA). The Cdc13-GFP signals (strong in the nucleus) faded in all of the anaphase cells (arrow) examined in scn1-17, as well as in wild-type cells. Bars, 10 µm. (C) The wild-type and scn1-17 mutant cells carrying plasmid pSAD1-GFP (Sad1 is a SPB protein) were observed for the spindle dynamics in living cells. Cells were cultured at 20°C for 9 hours. The white number in each panel shows the time (minutes) after starting the observations using a confocal micr oscope. Full anaphase spindle extension was complete within 40 minutes in wild-type cells, whereas spindle elongation was slow and incomplete after 96 minutes in scn1-17 mutant cells. Furthermore, the nuclear envelope in the middle appeared to remain in late anaphase, suggesting that nuclear division was not finished. Bars, 10 µm. (D) Distances between the SPBs were measured in living cells. In the scn1 mutant cells, the rate of anaphase spindle elongation was strikingly low and the SPBs did not reach the opposite poles of the cells, even after more than 100 minutes.
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