Fig. 4.

SMLM imaging of CENP-A with its chaperone HJURP (U2OS cells). a Wide-field images of CENP-A and HJURP show their overall co-localization at centromeres. b SMLM imaging demonstrates that CENP-A (red) and HJURP (green) form clusters of different shapes in which CENP-A and HJURP do not co-localize at the individual molecule level. Bottom left panels 1–8: magnified centromeres from image (b) demonstrate that an HJURP cluster is primarily situated in the empty center of a CENP-A cluster. The dSTORM images (panels b and 1–8) are reconstructed in the histogram mode with a bin size of 20 nm. The green rectangle in panel (b) indicates the region that is zoomed in and displayed as a Voronoi diagram in Supplementary Fig. 7. Scale bars, 2 µm (a), 500 nm (b), and 100 nm (1–8); for similar experiments using STED imaging see Supplementary Fig. 8. c, d Radial profiles of the rotationally averaged images of the CENP-A and HJURP particles at 1.5 h (c) and 5 h (d) after mitosis, using five cells for each time point; see Source Data file. e Schematic description of the evolution of the shape of CENP-A clusters during the cell cycle