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. 2008 Dec 1;183(5):819–834. doi: 10.1083/jcb.200804114

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© 2008 Whyte et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Mitotic localization of phosphorylated dynein. (1–4) NRK2 cells stained for chromatin (DAPI, blue), tubulin (green), and PT89-dynein (red) reveal PT89 at kinetochores at early prometaphase (1), late prometaphase (2), and on kinetochores of monooriented chromosomes (4). PT89-dynein is absent on kinetochores of aligned chromosomes at metaphase (3). The inset in panel 4 shows the image at a higher magnification. (5–8) Nocodazole-treated NRK2 cells were stained for chromatin (5 and 8), 74.1 antibody against the dynein ICs (6 and 8; Dillman et al., 1996), and PT89-dynein (7 and 8). Merged image confirms colocalization on kinetochores (8). (9–12) NRK2 cells stained for chromatin (9), PT89-dynein (10), and BubR1 (11) reveal colocalization of PT89-dynein with BubR1. (13–16) NRK2 cells stained for chromatin (DAPI, blue), tubulin (green), and PT89-dynein (13 and 14) reveal that the PT89-dynein signal is prominent on kinetochores of unaligned chromosomes but reduced on kinetochores close to the metaphase plate. V3 (against total dynein ICs; 15 and 16) is prominent on kinetochores during prometaphase but weaker along spindle fibers and spindle poles. In contrast, the V3 signal is weak on aligned kinetochores at metaphase but prominent along spindle fibers and at spindle poles (16, inset). Bars, 5 μm.