George von Dassow, Koen J.C. Verbrugghe, Ann L. Miller, Jenny R. Sider, William M. Bement (2011) CIL:15788, Strongylocentrotus purpuratus. CIL. Dataset

16-cell purple urchin embryo expressing three tandem GFPs fused to the ensconsin microtubule-binding domain (EMTB-3G; Faire et al., 1999). The embryo was treated with 20 µM trichostatin A (TSA), an inhibitor of HDAC6 (a tubulin deacetylase) that selectively disrupts dynamic microtubules in mammalian cells, at time 00:00. Within minutes of TSA addition in metaphase, asters were reduced to nearly nothing (04:00). Despite reduction in spindle length, cells enter anaphase (07:20) and complete cytokinesis (11:20–16:10). No microtubules connecting the mitotic apparatus to the cortex are visible. 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 at room temperature. Corresponds to Figure 4A and first segment of video 4 in J Cell Biol. 2009 Dec 14;187(6):831-45.

16细胞紫海胆胚胎表达三个串联融合绿色荧光蛋白（GFP），这些蛋白与微管结合蛋白域（EMTB-3G；Faire 等人，1999年）相连。该胚胎在00:00时接受了20 µM的三环素A（TSA）处理，TSA是一种抑制HDAC6（一种微管脱乙酰化酶）的抑制剂，它能选择性地破坏哺乳细胞中的动态微管。在TSA加入中期后不久，星体几乎消失（04:00）。尽管纺锤体长度减少，细胞于07:20进入有丝分裂后期，并在11:20至16:10之间完成细胞分裂。连接有丝分裂装置与细胞质膜的微管不可见。胚胎使用激光扫描共聚焦显微镜（Radiance 2000；Bio-Rad 实验室）进行成像，该显微镜安装在复合显微镜（E800；尼康）上，并使用40× 1.3 NA Plan-Fluor 油镜头在室温下进行成像。与《细胞生物学》2009年12月14日第187卷第6期的图4A和视频4的第一部分相对应。