Figure 9. BubR1 deletion constructs extend nocodazole arrest and positively impact error attachment machinery.
(A) Analysis of the time from nuclear envelope breakdown (NEBD) to anaphase onset in H2B-RFP wild-type MEFs infected with the indicated constructs with and without the addition of doxycycline (Dox) by live cell time-lapse imaging. n = 1 line, ≥ 20 cells per treatment. Data are mean ± s.e.m. ***p<0.001. FL, full-length. (B) In a nocodazole challenge, H2B-RFP wild-type MEFs infected with the indicated constructs with and without the addition of Dox were treated with 100 ng/ml of nocodazole and monitored by live cell time-lapse imaging. The point of time in which 50% of cells are arrested in mitosis is plotted. n = 1 line, ≥ 15 cells per treatment. *p<0.05, **p<0.01, ***p<0.001. (C) Analysis of chromosome misalignment in wild-type MEFs infected with the indicated constructs with and without addition of Dox. MEFs were treated with 100 µM monastrol for 1 hr and then with monastrol and 10 µM MG132 for 1 hr and released for 90 min into MG132. Cells were treated with DMSO (Vehicle) or 50 nM AZD1152-HQPA (AZD1152) throughout the duration of the experiment. n = 3 lines, 50 cells per line per treatment. Data are mean ± s.d. *p<0.05. (See associated Figure 9—source data 1).
DOI: http://dx.doi.org/10.7554/eLife.16620.028
Figure 9—figure supplement 1. Combined loss of Phe, D-box2 and KARD does not impact mitotic timing.
