Stabilization of microtubule dynamics suppresses detachment events of Dam1 phosphorylated kinetochores. (A) Western blotting to detect phosphorylation of the Dam1 complex in yeast cells. Dam1 detected with α-Dam1 antibody from cell lysates of log phase culture wild-type cells (black), untreated Cin8-Degron cells (blue), and Cin8-Degron cells treated with 30 µg/ml benomyl (magenta). (B) Quantification of phosphorylation levels from Western blotting. Each point represents an independent repeat. (C) Top: Representative image of an untreated phosphomimetic dam1-3D cell with attached kinetochores. Bottom: Representative image of a benomyl-treated phosphomimetic dam1-3D cell with attached kinetochores. (D) Tension distribution of untreated (n = 88 cells) and benomyl-treated (n = 94 cells) phosphomimetic dam1-3D cells, measured in population sampling experiments. (E) Fraction of dam1-3D phosphomimetic cells with detached kinetochores in untreated cells (purple, n = 261 cells) and in benomyl-treated cells (light pink, n = 226 cells; P << 0.00001, Z test), measured in population sampling experiments. (F) Top: Representative image of untreated wild-type cell with attached kinetochores. Bottom: Representative image of a benomyl-treated wild-type cell with attached kinetochores. (G) Tension distribution of untreated (n = 73 cells) and benomyl-treated (n = 64 cells) wild-type cells, measured in population sampling experiments. (H) Fraction of wild-type cells with detached kinetochores in untreated cells (black, n = 261 cells) and in benomyl-treated cells (gray, n = 226 cells), measured in population sampling experiments (P = 0.857, Z test; y-axis scaled to match panel 4 E). Inset: y-axis rescaled to demonstrate comparison between untreated and benomyl-treated cells. Source data are available for this figure: SourceData F4.