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. 2014 Apr 16;289(22):15463–15473. doi: 10.1074/jbc.M114.561019

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FIGURE 4. — Effect of unga knockdown or overexpression on the global DNA methylation level during early embryogenesis. A, B, D, and E, immunodetection of 5mC and H3 in nuclei (A and D) and the 5mC/H3 ratio (RGDM) (B and E). Embryos were injected at the one-cell stage with 10 ng of cMO, 10 ng of unga-MO, 500 pg of mcherry (mch) or unga-mcherry (unga-mch) mRNA and fixed at indicated stages for immunostaining. The RGDM was averaged from multiple nuclei and three to five embryos for each treatment. Ne, number of measured embryos; Nc, number of measured nuclei. **, p < 0.05; ***, p < 0.01. Error bars, S.D. C and F, detection of 5mC level by dot blotting using genomic DNA isolated from 256-cell stage embryos and anti-5mC antibody.