AIR-2 localizes normally in knl-1,3(kd) bivalents. AID triple depletions of KNL-1,3 and chromokinesin KLP-19 or SUMO E3 ligase GEI-17. (A–C) Representative time-lapse sequences of GFP-tagged AIR-2 (A and B) or mCherry-tagged SUMO (SMO-1; C) in control (A and C) or knl-3(kd) (B) embryos. Brackets in B indicate an intact bivalent segregating. (D) Time-lapse images of a spindle following overnight auxin-induced degradation of KNL-1,3, and KLP-19. (E) Time-lapse images of a spindle following overnight auxin-induced degradation of KNL-1,3, and GEI-17 displaying an abnormal disorganized structure. (F and G) No significant difference is observed in anaphase A or B velocities for KNL-1,3 double depletion or either triple depletion. (H) In 2/14 live time-lapse sequences, an intact bivalent (arrowhead) escaped the spindle in KNL-1,3 and KLP-19–depleted spindles during late anaphase I. It did not reincorporate into the MII spindle. Error bars and values are mean ± SEM. *, P < 0.05; ****, P < 0.0001. Bars = 3 µm. N, number of embryos; n, number of chromosomes. mCh MCH, mCherry; TUB, tubulin.