Figure 1.

NUSAP1 is a cell cycle-regulated microtubule-binding protein. A, U2OS cells were synchronized by overnight treatment with nocodazole and released by mitotic shake-off. Samples were analyzed by immunoblot as cells progress through the cell cycle. B, NUSAP1 localization to the mitotic spindle analyzing by immunofluorescent imaging of mitosis in U2OS cells. (Scale bars = 10 μm.) C, NUSAP1 localization was analyzed in nocodazole-treated cells and following incubation with ice-cold buffer to destabilize non-kinetochore microtubules. (Scale bars indicate 5 μm.) D, single plane confocal imaging of NUSAP1 localization on the spindle during metaphase. Insets highlight two kinetochore-microtubule attachments. (Scale bars indicate 5 μm.)