Skip to main content
. 2007 Apr 4;2(4):e342. doi: 10.1371/journal.pone.0000342

Figure 7. Over-expression of Mad3p induces bi-orientation and chromosome segregation defects.

Figure 7

A) Mad3p overexpression induces chromosome mis-segregation following checkpoint “challenge”. Strains were pre-synchronised with α-factor, released and then arrested with nocodazole. The nocodazole was then washed out and cells arrested in the next G1. Cells were fixed and analysed for the presence of GFP-marked chromosomes. Chromosome V was labelled with GFP and spindle poles with SPC42-mCherry. Arrowheads mark G1 cells containing two copies of chromosome V. B) Mad3p overexpression induces chromosome bi-orientation defects. Cells were pre-synchronised with α-factor, then depleted for Cdc20p through the addition of Methionine. Cells were then released from G1 into media containing nocodazole and benomyl. The microtubule drugs were then washed out to allow soindle assembly, but Cdc20p remained represed to arrest cells in metaphase. Cells were briefly fixed and scored for bi-orientation (breathing centromeres). Centromere IV was labelled with GFP and spindle poles with SPC42-tomato. Scale bars are 5 microns.