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. 2013 May 15;24(10):1559–1573. doi: 10.1091/mbc.E12-12-0850

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© 2013 Mitchison 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 1: — Pineapple assembly. (A) Typical field 1 h after addition of 5 μM Taxol and 5% DMSO to meiotic HSS containing tubulin (green) and NUMA (red) probes; 20× wide-field imaging. Note formation of similar assemblies across the whole field, comprising paired lines of microtubule bundles with NUMA aligned on the outside. (B) Round assemblies with open centers resembled a slice of canned pineapple (∼2 h of assembly, 20× wide field). Tubulin (green), NuMA (red). (C) A 40× confocal image at 90 min showing localization of Aurora B (red) on the inside and NUMA (blue) on the outside. Note global alignment of both types of foci on curved planes, which appear as lines in optical sections. (D) A 100× confocal image taken at 90 min showing one side of an open assembly. NUMA (blue) was on the outside as usual. Both NUMA and Aurora B (red) accumulate at foci. Aurora B foci are smaller and more numerous. (E) Pineapple assembly in C-HSS, which is completely free of organelles; 120 min, 20× wide field. Tubulin (green), NUMA (red). (F) Pineapple assembly in solution. This image was taken by assembly in solution for 1 h, followed by 20× dilution into a Taxol-containing buffer without DMSO, squashing between a slide and coverslip, and imaging within ∼5 min (40× confocal). Other examples are shown in Figure 4. NUMA (blue), Aurora B (red), tubulin (green).