FIGURE 2:

High K-MT stability correlates with efficient bipolar spindle maintenance without Eg5. (A–D) Images of BJ (A), RPE-1 (B), c33A (C), and HeLa (D) cells that were arrested in mitosis with 5 μM MG-132 for 90 min and then treated with 5 μM nocodazole for 6 min, extracted for 20 s, and fixed. Tubulin (green), centrin (yellow), and kinetochores (magenta; CREST) were detected by immunostaining. DNA (blue) was counterstained with Hoechst 33342. Dotted line shows region enlarged threefold. Scale bar, 5 μm. A and C are from a single experiment, and B and D are from a single experiment. Lookup tables (LUTs) were adjusted identically for images corresponding to the same experiment. (E) Quantification of residual MT polymer levels after nocodazole treatment. Puncta or short streaks were scored as “low polymer,” and long streaks joined at nodes were scored as “high polymer”; long streaks joined in two poles were scored as “spindle-like.” Data represent the mean ± SEM; n ≥ 100 cells from at least three experiments. *p < 0.001.