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. 2020 Apr 1;219(6):e201907082. doi: 10.1083/jcb.201907082

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© 2020 Jackman et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 3. — MAD1 binding to Cyclin B1 is required for genomic stability. (A) Chromosome number per cell was assayed for parental RPE yclin B1-Venus^+/−:Ruby-MAD2^+/− cells and the MAD1 E53/E56K clones 7D2 and 8B12 by metaphase spreads in three independent experiments. Black dots indicate 46 chromosomes and n indicates the number of cells assayed. Parental n = 89 cells, clone 7D2 n = 74 cells, clone 8B12 n = 80 cells. (B) The time from NEBD to anaphase was measured for parental RPE cyclin B1-Venus^+/−:Ruby-MAD2^+/− cells and the MAD1 E53/E56K clones 7D2 and 8B12 by time-lapse DIC microscopy. Violin plots show the data from three experiments (median time indicated by the black bar); n indicates the total number of cells analyzed. Parental n = 198 cells, clone 7D2 n = 197 cells, clone 8B12 n = 201 cells. (C) The duration of the mitotic arrest for parental RPE cyclin B1-Venus^+/−:Ruby-MAD2^+/− cells and the MAD1 E53/E56K clones 7D2 and 8B12 was assayed by time-lapse DIC microscopy in 55 nM nocodazole. Data are plotted as for B; n indicates the total number of cells analyzed. Parental n = 273 cells, clone 7D2 n = 228 cells, clone 8B12 n = 231 cells. The two-tailed P values were calculated using a Mann–Whitney unpaired t test.