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. 2015 Jan 15;26(2):256–269. doi: 10.1091/mbc.E14-10-1451

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© 2015 Ren et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — Spa2 colocalizes with Cdc15 and Imp2 in cytokinetic rings. (A) Representative live- cell images of the spa2-GFP cdc15-mCherry sid4-RFP at different cell cycle stages. Scale bar, 2 μm. (B) Arrested spa2-GFP cdc15-mCherry nda3-KM311 cells were fixed in ice-cold ethanol and then imaged. Scale bar, 5 μm. (C) Representative spa2-GFP cdc15-mCherry CR in initial stages of constriction rotated 90° and enlarged. (D) Images at 4-min intervals from a representative movie for spa2-mCherry₃ imp2-GFP sid4-RFP. Scale bars, 5 μm. (E) Time line showing the detection of various proteins at the division site. Time 0 was defined as SPB separation, and the mean time of detection of each protein ± SD is plotted. The onset of anaphase B was determined in n = 16 cells to be 12.7 ± 2.0 min, and maximum SPB separation (end of anaphase B) was at 26.7 ± 1.9 min, determined with n = 14 cells. Fic1 joined the CR at 9.6 ± 2.2, Cyk3 at 8.8 ± 1.6, Pos1 at 24.7 ± 2.7, and Pxl1 at 17.6 ± 3.6 min. n = 5 for Cdc15; 12 for Cyk3; 9 for Fic1; 9 for Pxl1; 8 for Rgf3; 11 for Imp2; 8 for Spa2; and 6 for Pos1.