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. 2010 Dec 27;191(7):1251–1260. doi: 10.1083/jcb.201005065

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© 2010 Breuer et al.

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Figure 5. — HURP is required for clustering extra centrosomes in U2OS cells. (A) HURP RNAi induces defects in clustering of extra centrosomes in cancer cells. PLK4-overexpressing U2OS cells (see Materials and methods) treated with control (luciferase) or HURP RNAi and stained for α-tubulin (red), centrin (green), and DNA (blue). (top) Control cells showing transient multipolar spindles during metaphase (left), and bipolar spindles with clustered centrosomes during anaphase/telophase (middle and right). (bottom) HURP-depleted cells showing multipolar anaphase/telophase figures. Insets show enlarged views of clustered centrosomes. (B) Knockdown of HURP in U2OS cells after 3 d of RNAi shown by Western blotting. (C) Multipolar fragmentary divisions in cells with extra centrosomes after HURP depletion. U2OS cells expressing GFP-H2B after control or HURP RNAi. (top) Control cell dividing bipolar. (bottom) Cell depleted of HURP dividing in a tripolar configuration of chromosomes before anaphase (inset). Time is shown as hours and minutes from chromatin condensation as t = 0. (D) Quantification of centrosome clustering defects in cells shown in C (***, P = 0.002). Error bars represent standard deviation. Bars, 10 µm.