Figure 4.

The major microtubule binding complex is in the distal brush tips. A) A cartoon schematic showing two possibilities for the location of Ndc80c. B) A graph showing the percent of kinetochores with at least one associated α-V5 gold particle relative to the total number of kinetochores. Kinetochores were purified from a strain with Ndc80-V5 epitope-tagged (Ndc80-V5, SBY21752; mean = 66%; st. dev = 3.3%) or not tagged (neg. control, SBY18150; mean=25%; st. dev=11%) and gold labeling was performed. Both groups contain 3 biological replicates. Error bars represent the standard error of the mean and white dots represent individual data points. C) Representative micrographs from gold labeling experiments targeting Ndc80-V5 kinetochores. Kinetochores are broken into sections as in (A). White arrows indicate a gold label in contact with a kinetochore. Scale bars are 200 nm. D) Quantification of the average amount of gold labels found in each section of the kinetochore as illustrated in (A) across 3 biological replicates. Individual data points are represented by dots and error bars represent the standard error of the mean. Blue indicates the Ndc80-V5 sample while green indicates the negative control which contains no V5 tag.