Figure 3
Ataxin-10-pS12 associates with the midbody.
(a–b) HeLa cells were stained with anti-pS12 (a) (green) or anti-Ataxin-10 antibodies (b), anti-α-tubulin antibodies (red) and DAPI. The arrow head pointed to the midbody. (c) The midbody-specific pS12 staining was dependent on Aurora B, but not on Plk1. HeLa cells were enriched in cytokinetic phase, then treated with 1 μm BI or 4 mM ZM, before staining with pS12 antibodies. Open arrowheads indicated staining at the midbody, while open arrows indicated absence of staining at the midbody. (d) The results were quantitated. Data represent mean ± SD of three independent experiments and more than 200 cells were counted in each experiment. Asterisks indicate significant differences from control cells, as determined by t-test (p1–2 = 0.001). Midbodies in a–d are maginifed and shown in insets. (e) The midbody-specific pS77 staining was dependent on Plk1 and Aurora B. Cells treated the same way as in (c) and then stained with pS77 antibodies and quantitated in (f). The p values are p4–5 = 0.004, p4–6 = 0.0045. Scale bar, 10 μm.
