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. 2009 Aug 28;4(8):e6831. doi: 10.1371/journal.pone.0006831

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This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.

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A) CENP-A levels are not significantly decreased in condensin-depleted cells. CENP-A levels were assessed in two independent samples of control and SMC2-depleted cells (both nocodazole-arrested as in Fig. 2E, 3 hr timepoint). Immunoblotting for CENP-A and RAN (as a loading control), quantified with Li-Cor Odyssey scanner using alternative fluorescent dyes. B) Schematic of cell synchronization, RNAi and SNAP-labeling strategy. CENP-A-SNAP synthesized during S phase was specifically fluorescently labeled in late S/G2 and the accumulation at centromeres was measured after mitosis at the end of the next G1. C) Results of procedure in (B). CENP-SNAP assembly at centromeres following indicated RNAi shown by TMR-Star labeling. Centromeres are counterstained with anti-CENP-C. D) Fluorescent intensity distribution of peak TMR-Star signal per cell from two independent experiments. The means and the standard errors of the mean (SEM) are shown for each experiment. The differences between the control RNAi and condensin depletion were statistically significant (one-tailed P-values<0.0001 in T-test with significantly different variances; t = 9.04 df = 740 in exp.1, t = 5.83 df = 1077 in exp.2). Eleven data points above 2000 were counted but are not shown on the graph.