Figure 4
Chromosome misalignment and abnormal kMT attachment are not repaired in Zwint-1-knockdown oocytes.
(a) Oocytes injected with either Zwint-1 dsRNA (dsZwint) or anti-Mad2 antibodies (Mad2 Abs) were treated with 25 μM MG132 at 4 h after GVBD. After 6 h culture, oocytes were cold-treated and immunostained with anti-tubulin antibodies, ACA and DAPI. The representative images from two independent experiments are shown. The area outlined in the white line is enlarged in the right panel. Scale bar, 10 μm. (b) Abnormal kMT attachments were scored. Data are mean ± SEM from two independent experiments (*p = 0.0002; **p < 0.005). The number of oocytes analyzed is shown above the bars. (c) Oocytes injected with either Zwint-1 dsRNA (dsZwint) or anti-Mad2 antibodies (Mad2 Abs) were cultured with 400 nM nocodazole from 4 h after GVBD and the PBE rate was determined at 13 h after GVBD. Data are mean ± SEM from three independent experiments (*p < 0.001). The number of oocytes analyzed is shown above the bars. (d) Oocytes were treated with (+) or without (−) MG132 at 4 h after GVBD and cultured for 6 h. Oocytes were matured to MetII stage by releasing from MG132-mediated inhibition and cultured with monastrol for 2 h prior to chromosome spreads. Arrows indicate the unpaired chromatids. Scale bar, 10 μm. (e) Quantification of aneuploidy. Data are obtained from three independent experiments with the indicated number of oocytes shown above the bars. (*p = 0.007; ns, not significant).
