Fig 1.
Schematic structure of studied imprinted genes on mouse chromosome 7q.
Proximally, near the centromere, are Paternally expressed gene 3 (Peg3) and Small Nuclear Ribonucleoprotein Polypeptide N (Snrpn), and distally Insulin-like growth factor 2/H19 locus (Igf2/H19). Gray boxes illustrate maternally or paternally expressed active alleles and black boxes inactive alleles. Imprinting control regions (ICR) or differentially methylated regions (DMR) are marked with methylated (black) or unmethylated (white) balls. Arrows and sequences below the ICRs or DMR represent the regions of interest. Not drawn into scale.
Fig 2.
DNA methylation levels of CpG sites at the Igf2/H19 ICR (A), Igf2 DMR1 (B), Snrpn ICR (C), Peg3 ICR (D), and Line-1 (E) in 9.5 embryonic days old (E9.5) control and alcohol-exposed embryos and placentas. Control samples are colored in black and alcohol-exposed samples in gray. The numbers of samples are presented in columns. CpG sites 3 and 5 at Igf2 DMR as well as CpG site 13 at Peg3 ICR in E9.5 placentas have been excluded since no methylation value could be detected by EpiTYPER. Error bars denote the SD.
Fig 3.
Expression levels of imprinted genes in control and alcohol-exposed 9.5 embryonic day old (E9.5) embryos and placentas as well as E16.5 placentas.
Igf2, H19, Snrpn and Peg3 expressions in control and alcohol-exposed E9.5 embryos (A), E9.5 placentas (B) and in E16.5 placentas (C). Control samples are colored in black and alcohol-exposed samples in gray. The numbers of samples are presented in columns (expression of Snrpn in one embryo and one E9.5 placenta samples was not detected). Error bars denote the SD. P-value; (A) two-way Student’s t-test: *p<0.05, **p<0.001, (C) Two-way ANOVA: *p<0.05.