Figure 6.

Hec1 localizes to the kinetochore outer plate. HeLa cells grown to 60-80% confluency were treated with 20 μM nocodazole for 3 h, extracted, and fixed. Fixed cells were treated with monoclonal anti-Hec1 primary antibody and anti-mouse secondary conjugated to 0.8-nm gold clusters that were then enhanced with silver reagent. (A) Low-magnification overview. Linear concentrations of silver-enhanced gold particles distinguishes kinetochore labeling (arrows) from the sparse occurrence of particles in the background. (B-D) Higher magnification examples of kinetochore labeling. Arrows indicate regions where the outer plate is visible underneath gold particles. (E) Enface or semienface views of labeled kinetochores (arrows). Note the more circular arrangement of gold particles. Scale bars: (A) 1 μm; (B-E) 200 nm.