Schematic of the in vitro kinetochore assembly assay. Individual Atto565-labeled centromeric DNAs were tethered sparsely onto a PEG passivated coverslip surface through biotin-avidin linkages. The surface-tethered DNAs were then incubated for 60 min with yeast whole-cell lysate derived from strains with GFP-tagged kinetochore components (Ndc10, Cse4, Ctf19, or Ndc80). Kinetochores assembled spontaneously onto the centromeric DNAs and were then imaged with TIRF microscopy after washing out the extract. (coverslip. Side-captured microtubules mainly rotated in a two-dimensional (2D) plane parallel to the coverslip in a propeller-like fashion. Larson JD, Heitkamp NA, Murray LE, Popchock AR, Biggins S, Asbury CL. Kinetochores grip microtubules with directionally asymmetric strength. J Cell Biol. 2025 224(1):e202405176. PMID: 39485274