Fig. 4: Sub-classification of centromeres by cohesin localization.

a A representative centromere with CENP-A (cyan) and RAD21 (magenta) signals and the line profile (gray) used for measurements is shown. Center-enriched RAD21 ratio was determined by measuring the ratio of intensity at pixels (white) at the center of the centromere and 6 pixels vertically away from the center. b Chromosomes were assigned ranks based on center-enriched RAD21 ratio (Ratio 0.0861—Rank 1, Ratio 5.519—Rank 463). A montage is shown in which the x axis represents each centromere, and the y axis represents the vertical profile through the middle of the centromere region, sorted by their rank. Three classes—split (ranks: 1–200), uniform (ranks:201–350) and central (ranks:351–463) were arbitrarily assigned based on these ratios. c Aligned and averaged images for each class are shown. x and y axis on images indicate distance (nm). d Average intensities of RAD21 along the vertical profile in the three classes are shown. e Inter-CENP-A distances measured for the three classes—central (n = 200), uniform (n = 150), split (n = 113) are compared by unpaired one-way Kruskal-Wallis test (multiple comparisons). Dotted white lines in violin plots denote median values. n.s. - not significant.