Z-focal series HeLa cell at metaphase stained for DNA (blue), Aurora B (green) and phospho-Ser10-histone H3(red) after depletion of Bod-1, a protein that associates wtih a large macromolecular complex and localizes with kinetochores and spindle poles during mitosis. This control for this Bod-1 depletion experiment is CIL: 13375.
Cells were fixed with 3.7% paraformaldehyde or methanol for 2 min at −20°C. Mouse anti–α-tubulin DM1A (Sigma-Aldrich), and mouse anti–Aurora B antibody AIM-1 (BD Biosciences) were used at 1:500. Fluorescently labeled secondary antibodies were all obtained from Jackson ImmunoResearch Laboratories. Image corresponds to Figure 5A from JCB vol. 179 no. 2 187-197.
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