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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 6. — Spartin is not required for Ist1 localization to midbodies. (A) HeLa cells were transfected with control siRNA or else spartin-specific siRNA and immunostained for spartin (green) or β-tubulin (red). Boxed areas are enlarged in the panels below. Merged images are shown at the right. (B) Cells treated with spartin siRNA were immunostained for Ist1 (green) and β-tubulin (red). Ist1 is present at the midbody, as shown in the boxed area enlarged in the panels below. A DIC image centered on the midbody is shown in the inset. (C) Cell extracts from HeLa cells treated with control or spartin-specific siRNA were immunoblotted for spartin. GAPDH levels were monitored to control for protein loading (left). Quantification of the percentage of control and Ist1 siRNA cells with spartin (middle) or Ist1 (right) present at midbodies (means ± SD of at least three trials, with 100 cells per experiment). (D–G) Quantifications of cells prepared as described in C are shown for percentages of mitotic cells (D), cells in interphase (E), cells interconnected with a visible midbody (F), and multinucleated cells (G). Means ± SD of at least three trials are shown, with 100 cells per experiment. **p < 0.001. Bars, 10 μm.