Single-plane sequences of anucleate, centrosome-containing cytoplasts (sand dollars) expressing three tandem GFPs fused to the ensconsin microtubule-binding domain (EMTB-3G; Faire et al., 1999). This anucleate cytoplast has four centrosomes, two in focus. In normal cells this size, furrowing would begin between 03:00 and 07:00, but no furrow occurs even 20 min later. Centrosomes are closer than the distance to the equator. Embryos were imaged with a laser scanning confocal microscope (Radiance 2000; Bio-Rad Laboratories) mounted on a compound microscope (E800; Nikon) using a 40× 1.3 NA Plan-Fluor oil lens. Room temperature was maintained at 14–17°C. Video corresponds to the first segment of video 6 and Fig 7A in J Cell Biol. 2009 Dec 14;187(6):831-45.
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