Figure EV5. Chromosome segregation defects in DNA2 null cells (see also Fig 4).

• A
  Attachment of microtubules to the centromeres in WT and DNA2 null cells. Shown are the original images used to generate the merged images in Fig 4A. Movies of the 3D rotations of projections of these cells are shown on the right. Scale bars, 2 μm.
• B, C
  Methodology for lagging chromosome (B) and congression index analysis (C). A lagging chromosome is defined as a cluster of DNA lacking anti‐CREST and anti‐CENP‐A staining, as well as lack of attachment to α‐tubulin (white arrows). The congression index was calculated as the ratio of metaphase chromosome width (parallel to the spindle poles) to length (perpendicular to the spindle poles). Scale bars, 2 μm.
• D, E
  Microscopic analysis of chromosome segregation in the DNA2^Flox/+/+ and DNA2^Flox/−/− cells. Panel (D) shows representative anaphase cells, which were analyzed by staining with DAPI (blue) and anti‐CENP‐A (red). Panel (E) shows the percentage of cells with segregation abnormalities (mean ± SD of three biological repeats, > 100 cells for each group). *P‐value is < 0.05 using unpaired two‐tailed t‐test. Scale bars, 5 μm.