Fig. 2.

MLL5-depleted cells fail to align chromosomes on metaplate because of impaired function of the kinetochore-MT attachment error-correcting machinery. (A) Metaphase-arrested cells were categorized into three groups: bipolar normal alignment, bipolar misalignment and multipolar misalignment. Experiments were repeated three times and the average percentage is presented. Scale bars: 5 µm. (B) The kinetochore-MT capture and attachment process were examined in a cold-induced MT depolymerization assay. Kinetochore fibers were stained with α-tubulin antibody and kinetochores were labeled with CREST. Scale bars: 5 µm. (C) The average inter-kinetochore distances (for sister kinetochores) were measured, as shown, in control cells (n = 70) and MLL5-depleted cells (n = 84). Scale bars: 5 µm. (D) A time-course study of the chromosome alignment process was conducted in NC- and MLL5-depleted cells after monastrol release. Cells were categorized into four groups: monopolar, bipolar misalignment, multipolar misalignment and bipolar normal alignment, and the percentages for each group at each time point after monastrol release are presented (n = 100 cells per sample). Scale bars: 5 µm.