Figure 2
Mad2-mediated SAC inactivation in Zwint-1-knockdown oocytes.
(a) Oocytes injected with the indicated dsRNA were cultured with 400 nM nocodazole from 4 h after GVBD and the PBE rate was scored at 13 h after GVBD. Data are mean ± SEM from three independent experiments (*p < 0.0001). The number of oocytes analyzed is shown above the bars. The exogenous expression of human Zwint-1 (hZwint) was shown by immunostaining in the right panel. Scale bar, 10 μm. Note that the PBE induced by Zwint-1 knockdown was rescued by overexpression of hZwint. (b,c) Zwint-1-knockdown oocytes were fixed at 2–3 h (Pro-MetI) or 6 h (MetI) after GVBD and immunostained with either anti-BubR1 (b) or anti-Mad2 (c) antibodies. Control oocytes injected with dsEGFP were fixed at 4 h or 8 h after GVBD corresponding to Pro-MetI or MetI, respectively. Kinetochores and DNA were stained with ACA and DAPI, respectively. Scale bar, 10 μm. Quantification of fluorescent intensity of BubR1 and Mad2 is shown in the right panel of images. Data are mean ± SEM of the indicated number of oocytes from two independent experiments (*p < 0.0001).
