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Fig. 3 | Epigenetics & Chromatin

Fig. 3

From: Oxidative stress in sperm affects the epigenetic reprogramming in early embryonic development

Fig. 3

The BER machinery is sequestered to paternal pronuclei of zygotes derived from sperm exposed to oxidative stress. a γH2AX (green) levels were assessed by immunofluorescence of zygotes 20 h after IVF using γH2AX antibodies. Representative images of PN3/4 zygotes are shown. Quantification of γH2AX is represented as a ratio of the mean signal intensity between the two pronuclei signal (mean γH2AX intensities pPN/mPN). (n = 24 zygotes control and n = 17 H2O2-treated zygotes; experiment replicated three times independently.) Each data point represents the mean signal of the ratio (pPN/mPN) within an independent zygote. b Representative immunofluorescence images of PN3/4 zygotes 20 h after IVF stained with XRCC1 antibody (green) are shown. Quantifications of XRCC1 are shown as ratio of the mean signal intensity between the two pronuclei signal (pPN/mPN). Each data point represents the value of four independent experiments, using 120 zygotes for the control group and 85 zygotes from +H2O2 groups. Statistical analysis was performed using Student’s t-test (two-tailed). Error bars indicate s.d.; **P < 0.01; ns: non-significant and refers to P = 0.075. Maternal pronucleus: mPN/♀; paternal pronucleus: pPN/♂. Scale bars, 50 μm

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