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. 2010 Jul 12;190(1):35–43. doi: 10.1083/jcb.201004017

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© 2010 Rankin and Wordeman

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Figure 2. — Severe spindle rocking is driven by cortical myosin and long astral MTs. (A) Still images from Video 4 showing spindle oscillations in a HeLa cell expressing EGFP–myosin IIA (green) and mCherry–α-tubulin (red) and treated with MCAK-specific siRNA for 36 h. Asterisks mark the location of chromosomes. Time is relative to anaphase onset. (B) A schematic of kymographs in C is shown. The dashed box indicates the slice plane for the construction of the kymographs. d, distance; t, time. (C) Kymographs of cells in anaphase with normal (left), weak (middle), and severe (right) rocking. Arrows indicate bleb and subsequent movement of the spindle. (D) Astral MTs are longer in metaphase HeLa cells treated with MCAK-specific siRNA for 48 h (bottom) relative to control cells (top). MTs are labeled with anti–α-tubulin (green), actin filaments with Texas red phalloidin (red), and DNA with DAPI (blue). Insets show magnified views of boxed regions. (E) MT stability increases in interphase when MCAK is depleted, as indicated by staining with anti–Glu-tubulin antibodies (right) after treatment with control (top) and MCAK-specific siRNA (bottom) for 48 h.