Fig. 4

Reduced Net1 activates the SAC and increases the incidence of aneuploidy. (A) Control and Net1-KD oocytes were immunolabeled with anti-BubR1 antibody (green) and counterstained with PI to examine chromosomes (red). Representative confocal images of pre-MI and MI oocytes are shown. Arrowheads indicate scattered chromosomes in Net1-KD oocytes (scale bar, 2.5 μm). (B) Quantification of BubR1 fluorescence intensity in control and Net1-KD oocytes (19.3 ± 2.14% vs. 71.5 ± 1.48%). (C) Chromosomal spreads of control and Net1-KD MII oocytes (chromosomes were stained with Hoechst 33,342 [blue]). Representative confocal images show euploid control oocytes and aneuploid Net1-KD oocytes. Arrows indicate the premature separation of sister chromatids (scale bar, 2.5 μm). (D) Quantification of aneuploidy in control and Net1-KD oocytes (10.1 ± 1.33% vs. 30.9 ± 1.85%). Graph shows the mean ± SD of the results obtained from three independent experiments (*significantly different at P < 0.05, **significantly different at P < 0.01)