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. 2016 May 11;7(21):30155–30165. doi: 10.18632/oncotarget.9310

Figure 6. Interaction between Zfp207 and Bub3 in mouse oocytes.

Figure 6

A. Protein levels of SAC components in Zfp207-knockdown oocytes were determined by western blotting. The blots of control and Zfp207-MO injected oocytes were probed with anti-Zfp207, anti-Bub3, anti-BubR1, anti-Bub1, anti-Mad2, and anti-β-actin antibodies, respectively. B. Colocalization of Zfp207-mCherry and Bub3-GFP. GV oocytes were co-injected with cRNAs of Zfp207-mCherry and Bub3-GFP, and then cultured to Pro-MI stage. Signals of mCherry and GFP were acquired under confocal microscope at 594nm and 488nm laser, respectively. Chromosomes were counterstained with Hoechst. Scale bar, 5μm. C. Zfp207 associates with Bub3 in oocytes. Co-IP was performed to determine the interaction between Zfp207 and Bub3. Oocyte lysates were incubated with IgG and anti-Bub3 antibody, respectively, followed by incubation with protein G beads. The blots of IP eluates were probed with anti-Zfp207 and anti-Bub3 antibodies, respectively. D. Quantitative analysis of aneuploid eggs in the control, Zfp207-KD and Bub3-rescue oocytes. Data were presented as mean percentage (mean ± SEM) of at least three independent experiments. Asterisk denotes statistical difference at a p < 0.05 level of significance.