Distinct pole-unfocusing phenotype after Asp or Ncd depletion. (A–C) GFP-tubulin imaging during acentrosomal spindle formation after Asp or Ncd RNAi. Cells were depleted of centrosomes by Cnn RNAi. In the absence of Asp, poles were unfocused throughout prometaphase and metaphase (9–20 min in B), whereas in the absence of Ncd, most cells contained transient focused poles during prometaphase (arrowheads in C). Bar, 5 µm. (D) Change in pole width during prometaphase and metaphase of a representative cell after each RNAi. The first time point represents the timing when overall bipolar spindles were first detected (a few minutes after NEBD). Time 0, anaphase onset. Pole width was measured after z-stack projection. (E) Quantification of the pole phenotype. A focused score was applied when one or more focused poles were transiently or continuously observed during prometaphase.