Z-series of fluorescence images of a Hela cell at mitotic metaphase stained for DNA (blue), Aurora B (green) and tubulin (red). This image is the control for the Bod-1 depletion experiment illustrated in CIL: 13372. Bod-1 is a protein that associates with a large macromolecular complex and localizes with kinetochores and spindle poles during mitosis.
Cells were fixed with 3.7% paraformaldehyde or in 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.