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. 2022 Nov 18;33(14):br25. doi: 10.1091/mbc.E22-07-0297

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FIGURE 4: — Ring complexes are not properly removed from chromosomes in him-18 mutant anaphases. (A) Wild-type (EU1067) and him-18(tm2181) during Anaphase I; shown are DNA (blue) and tubulin (green). Anaphase defects were quantified using sum projections of the DNA channel; him-18(2181) mutants had frequent anaphase defects, as previously reported (Saito et al., 2009). (B) Wild-type (EU1067), akir-1(rj1), and him-18(tm2181) spindles arrested in early Anaphase I using mel-28(RNAi). Shown are DNA (blue), AIR-2 (red), tubulin (green, column 1), and SUMO (green, column 2). Single-slice zooms show DNA (magenta) and SUMO (green). While there is a space between SUMO and DNA in wild-type and akir-1(rj1) anaphases, indicating that RCs have disengaged from chromosomes, RCs in him-18(tm2181) remain associated with chromosomes, looping around the short-arm nubs. (C) Quantification of RC localization following mel-28(RNAi); the anaphase ratio was calculated by measuring the distance between the two farthest rings, and then dividing that distance by the pole-to-pole spindle length. RCs in him-18(tm2181) spindles occupy a larger fraction of the spindle than in wild-type anaphase (p = 0.004; two-tailed Student’s t test). Scale bars = 5 µm (full images), 1 µm (zooms).