Testing the effect of EDs on imprint establishment by assaying the allele-specific DNA methylation at DMRs in G2 embryos. (A)
In utero exposed male 129S1 G1 fetuses were grown to adulthood and mated with unexposed JF1 females; the resulting G2 fetuses (three blue stars) derived from exposed prospermatogonia. (B)
In utero exposed G1 females were mated with unexposed JF1 males to generate G2 offspring (three red stars) which derived from exposed primary oocytes. G2 fetuses were dissected at 13.5 dpc to collect organs. (C, D) MIRA-SNuPE results of 13.5 dpc G2 embryo DNA samples after potential paternal and maternal transmission of aberrant DNA methylation at DMRs. The heatmap shows the percentage of average parental allele-specific methylation in organs; the color scale is as in Figure 2C. Letters in parentheses indicate different SNPs. Statistically significant differences relative to control (oil) greater than 5% and 10% are indicated by thin or bold rectangles, respectively (Student’s t-test, P <0.05).