Kinetochore location of Zwint1, RZZ complex, Mad1, and Spindly in metaphase HeLa cells expressing Spindly containing the motif mutation (GFP-SpindlyF258A). (A, C, E, and F) Example red and green immunofluorescence image pairs used for Delta analysis. Bars, 5 µm. (A′–B′, C′, D, E′, E″, and F′) Example plots of measured Delta values as a function of interkinetochore (K-K) stretch. (G) Comparison of distances measured between CENP-I and three different epitopes in control and Spindly motif mutant metaphase cells. The Hec1–CENP-I separation measurements are replotted from Gassmann et al. (2010). Error bars are SD from the means. (H) Images of a typical sister kinetochore pair for each of the epitopes whose centroids have been measured (indicated by the labels at the bottom) using Delta analysis. Bar, 1 µm.
