NOD chimeras with either plus end–directed motility or tip tracking activity produce PEFs. (A) Distance between the monopole center and Ndc80-stained kinetochores. The distance between kinetochores and monopole centers decreased by 30% in the absence of NOD and was rescued in NOD-depleted cells by inducing either EB1-NOD-mCherry or kinesin-1-NOD-mCherry (control, n = 283 kinetochore pairs; NOD RNAi, n = 302; NOD RNAi + EB1-NOD-mCherry, n = 293; NOD RNAi + kinesin-1-NOD-mCherry, n = 383). Error bars are SEM. (B) Representative maximum projection images for each condition. Monopoles were generated in each condition by depleting Klp61F. The chimeric NOD-mCherry protein is shown only in black and white. In the merged images DNA is blue, Ndc80 is red, and microtubules are green. (C) Histograms of the distribution of pole–kinetochore measurements for the four experimental conditions each fit with a Gaussian function (black lines). The dashed line extending through the histograms marks the mean pole–kinetochore distance in control RNAi-treated cells. Two-tailed p-values are shown. Bar, 10 µm.