Fig. 2. Single scn1 mutant is defective in spindle elongation, nuclear positioning and cell division, producing frequent late mitotic phenotypes at 20°C. (A) Wild-type (WT) and scn1-17 grown at 33°C were transferred to 20°C in the YPD medium. Anti-tubulin (red), anti-Sad1 (green) and DAPI (blue) staining were done for wild-type and scn1 mutant cells after 9 hours and 12 hours at 20°C. Late anaphase cells, revealing separated chromatids and a spindle that was not fully elongated, were marked as A. The nuclear displacement indicated as B was often seen in scn1 mutant cells. Cells lacking the nuclei were also seen (an example indicated as scn1-17 in right panel). Bar, 10 µm. (B) The frequency of late anaphase cells in the wild-type and scn1-17 mutant after the transfer to 20°C for 0-12 hours. Cells in late anaphase were significantly accumulated after 9 hours in scn1-17. (C) The frequencies of wild-type (upper) and scn1 mutant (lower) cells showing normal positioning of the separated daughter nuclei (open rectangles), abnormally displaced daughter nuclei (filled triangles) and no nuclei (crosses) are shown.