Original images used for preparation of figures in Guin et al., 2026, eLife

These files include original images obtained from confocal microscopy, PCR gel electrophoresis and Western blot experiments and used for preparation of figures in this article- Guin et al., "Orderly mitosis shapes interphase genome architecture." eLife, 2026, DOI: https://doi.org/10.7554/eLife.108410.1Abstract:Genomes assume a complex 3D architecture in the interphase cell nucleus. Yet, the molecular mechanisms that determine global genome architecture are only poorly understood. To identify mechanisms of higher order genome organization, we performed high-throughput imaging-based CRISPR knockout screens targeting 1064 genes encoding nuclear proteins in human cell lines. We assessed changes in the distribution of centromeres at single cell resolution as surrogate markers for global genome organization. The screens revealed multiple major regulators of spatial distribution of centromeres including components of the nucleolus, kinetochore, cohesins, condensins, and the nuclear pore complex. Alterations in centromere distribution required progression through the cell cycle and acute depletion of mitotic factors with distinct functions altered centromere distribution in the subsequent interphase. These results identify molecular determinants of spatial centromere organization, and they show that orderly progression through mitosis shapes interphase genome architecture.