knl-1,3(kd) metaphase spindles have chromosome congression and alignment defects. (A and B) Single time points from time-lapse sequences of metaphase spindles depleted of KNL-1 and KNL-3 by auxin-induced degradation (A) or GFP(RNAi) (B). Arrows indicate chromosomes on periphery of spindle. The bright green body in the last image is that of a nearby sperm. (C) The relative position of each bivalent along the pole-to-pole axis of its spindle. A congressed bivalent has an expected ratio of 0.5. (D) The angle of individual chromosomes relative to the pole-to-pole axis was determined in five control and nine knl-1,3(kd) metaphase spindles. The alignment of each chromosome is illustrated below the graph. (E) The percentage of knl-1,3(kd) spindles with chromosomes on the outside edge of the spindle. Error bars indicate mean ± SEM. Bars = 4 µm. ****, P < 0.0001. mNG, mNeonGreen; TUB, tubulin.