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. 2015 Jan 3;13(24):3828–3838. doi: 10.4161/15384101.2014.967066

Figure 4.

Figure 4.

Pre-MBT embryos can initiate a DNA damage response. (A) Embryos were injected with Alexa 594-Histone H1 protein and treated with 10 Gy IR as indicated, then imaged live during cleavage-stage cell cycles. Images (A) show cell cycle progression based on Histone H1, starting at ∼2.5 HPF (t = 0), with insets as in Figure. 1A. Insets are shown with higher contrast settings so nuclei morphology are clear. Cell cycle lengths were measured as time between M-phases, based on Histone H1 morphology, and averaged over cleavages 5-9 for multiple embryos (n ≥ 9). (B) Pre-MBT (1.5 HPF) or post-MBT (4 HPF) embryos were treated with 10 Gy IR then fixed and stained for γH2AX and DNA. γH2AX staining intensity was quantified and averaged over multiple embryos (n ≥ 10) for each condition. Similar results were obtained in 4 independent experiments; a representative plot is shown. Error bars indicate s.e.m., *P < 0.05. All scale bars 20 μm. Arrowheads show examples of nuclear fragmentation and lagging chromosomes from DNA damage in the irradiated embryo.