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. 2021 Feb 9;8:610773. doi: 10.3389/fcell.2020.610773

Figure 2.

Figure 2

Maternal UHRF1 depletion causes DNA hypomethylation at oocytes and zygotes. (A) The representative immunofluorescent-staining images for anti-5-methylcytosine (anti-5-mC) of control and Gdf9-cKO GV oocytes are shown. Nuclei were stained with DAPI. Scale bar = 50 μm. (B) Histogram shows a semi-quantitative analysis regarding changes of 5-mC distribution of fluorescence intensity at GV oocytes of (A). Quantification of the level of 5-mC in control and Gdf9-cKO oocytes and data are presented as means ± SEM from three independent experiments (total number of GV oocytes analyzed: n = 92 for control, n = 81 for Gdf9-cKO). *p < 0.05. (C) Representative immunofluorescent-staining images for 5-mC of control and Gdf9-cKO zygotes are shown. Nuclei were stained with DAPI. Scale bar = 10 μm. (D) Histogram shows a semi-quantitative analysis of changes in the fluorescence intensity, indicating the 5-mC distribution in zygotes of (C). Data are presented as mean ± SEM. n = 3 independent experiments (total number of zygotes analyzed: n = 131 for control, n = 73 for Gdf9-cKO). *p < 0.05. (E,F) Representative immunofluorescent staining images (E) and the quantification of fluorescence intensity (F) for 5-hydroxymethylcytosine (5-hmC) of control and Gdf9-cKO GV oocytes are shown. Data are presented as mean ± SEM. n = 3 independent experiments (total number of GV oocytes analyzed: n = 121 for control, n = 87 for Gdf9-cKO). (G,H) Representative immunofluorescent-staining images (G) and fluorescence intensity quantification (H) for 5-hmC of control and Gdf9-cKO zygotes are shown. Data are presented as mean ± SEM. n = 3 independent experiments (total number of zygotes analyzed: n = 95 for control, n = 78 for Gdf9-cKO). Scale bar = 50 μm. *p < 0.05.