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. 2009 Jun 1;185(5):779–786. doi: 10.1083/jcb.200812034

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© 2009 Dupin 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 4. — Actin and nucleus-dependent forces control centrosome positioning. (A–D) Astrocytes were plated onto fibronectin-printed micropatterns. After 4 h, cells were treated with 1 µM cytochalasin D (CytoD), 20 µM nocodazole (Noco), both (CytoD + Noco), or were enucleated and incubated in the presence (Enucleation + Noco) or absence of nocodazole (Enucleation) for another 3 h. (A) Pan-cadherin (green), pericentrin (red, white arrowhead), and Hoechst (blue) stainings. (insets) Higher magnification images of cadherin staining at cell–cell contacts. Bar, 20 µm. (B) Distances between the nucleus and cell centroids (black) and between centrosome and cell centroid (white). Data are given as mean ± SD of three independent experiments and a total of at least 120 cells. (C) Histograms showing the distribution of angles formed by the centrosome–nucleus axis and the micropattern radius passing through the centrosome. The median angle is indicated in red. Statistical differences between control and inhibitor-treated cells are indicated. Schematics depict the typical intracellular organization in each condition. (D) Pan-cadherin (green), anti-pericentrin (red, white arrowhead), and Hoechst (blue) stainings. Enucleated cells are indicated by an asterisk and nonenucleated cells are indicated by “n”. Bottom panels show higher magnification images of an enucleated cell. (E) Distance between the centrosome and the cell centroid in enucleated (“without nucleus”) and nonenucleated cells (“with nucleus”). Data are given as mean ± SD of three independent experiments, with a total of at least 100 cells. *, P < 0.05; **, P < 0.005.