z-focal series through Hela cell in metaphase illustrating MCAK localization in Bod-1siRNA cells. Cells were transfected with control (this image) or Bod1 (CIL: 13378) siRNA. After 72 h, cells were treated with monastrol for 3 h and released into media containing MG132 for 1 h before fixing. Cells were stained for total mitotic centromere-associated kinesin (MCAK) population. MCAK localizes to the inner centromere in its phosphorylated form but concentrates at kinetochores in its dephosphorylated state.
Antibodies were rabbit anti-HEC1 (green) antibody (Abcam) used at 1:1,000. and sheep anti-MCAK (red) (Andrews et al., 2004) used at 1 μg/ml, and DNA (blue). Fluorescently labeled secondary antibodies were all obtained from Jackson ImmunoResearch Laboratories. Corresponds to Fig 5B in JCB vol. 179 no. 2 187-197.
Spatial Axis | Image Size | Pixel Size |
---|---|---|
X | 355px | 0.0629µm |
Y | 381px | 0.0629µm |
Z | 28px | 0.2µm |