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. 2010 Oct 1;21(19):3293–3303. doi: 10.1091/mbc.E09-10-0879

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© 2010 by The American Society for Cell Biology

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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Figure 5. — Ist1 depletion decreases spartin recruitment to midbodies and impairs cell division. (A) HeLa cells were transfected with control siRNA or else Ist1 siRNA and immunostained for Ist1 (green) or β-tubulin (red). Boxed areas are enlarged in the panels below. Merged images are shown at the right. (B) Ist siRNA cells were imunostained for spartin (green) and β-tubulin (red). There is no spartin at the midbody, as shown in the boxed area enlarged in the panels below. (C) Cell extracts from HeLa cells treated with control or Ist1-specific siRNA were immunoblotted for Ist1. PLCγ levels were monitored to control for protein loading (left). Percentages of control and Ist1 siRNA cells with spartin (middle) or Ist1 (right) present at midbodies are shown (means ± SD of at least three trials, with 100 cells/experiment). (D and E) Multinucleated cells were evident upon Ist1 siRNA treatment of HeLa cells (D). Nuclei were identified by costaining with DAPI. (E) Quantification of the percentage of multinucleated cells after control versus Ist1 siRNA treatment (means ± SD of at least three trials, with 100 cells per experiment). *p < 0.001. Bars, 10 μm.