Fig. 4: Orientation-coupled removal of dynein from kinetochores in the kinetochore dynein module-only state.

a Snapshot of dynein localization, visualized using in situ-tagged DHC-1::GFP, at a time point when chromosomes are autonomously orienting on the spindle in the kinetochore dynein module-only state. Numbered circles highlight 3 chromosomes in different states. The genotype and RNAi conditions employed here and in (b–d) are shown below the image. Scale bar, 2 µm. n is the number of embryos analyzed. b Image sequence of a single chromosome and of dynein localized to its two sister kinetochores. To aid comparison, the panels are centered on the chromosome; dots (no movement) and arrows (directional movement) in each panel indicate chromosome movement relative to the spindle pole. Boxed region shows a subset of panels where the chromosome was not centered to highlight coupled poleward translocation and orientation associated with the capture of the right kinetochore. Green and brown shaded boxes highlight successive capture of the two sister kinetochores. Time is relative to anaphase onset in seconds. Graph on the lower right quantifies the DHC-1::GFP fluorescence intensity on each kinetochore, along with the chromosome angle relative to the spindle axis. Shaded areas on the graph correspond to the shaded boxes around image panel sets. Symbols below −54 s and −27 s panels serve as reference points linking image panels to the graph. Scale bars, 1 µm. c Analysis of kinetochore dynein fluorescence intensity and chromosome angle for 9 kinetochores. Shading in the circle indicates DHC-1::GFP fluorescence signal and the black line indicates the chromosome angle relative to the spindle axis. For simplicity, translocation on the spindle is not shown. d DHC-1::GFP localization on a chromosome that maintains a persistent lateral orientation through anaphase and does not biorient. Scale bar, 1 µm. n is the number of chromosomes analyzed. Source data are provided as a Source Data file.