Fig. 2
Prenatal exposure to E3 alters global gene expression, DNA methylation, pAKT protein levels, and E2 responsiveness in female offspring uteri. Eight-week-old pregnant female CD-1 mice were treated with vehicle DMSO (CT) or E3. At 8 weeks after birth, the female offspring uteri were analyzed. A, B Quantification of selected gene expression in uteri by quantitative RT-PCR. A The representative genes with decreased expression after fetal E3 treatment. B The representative genes with increased expression after fetal E3 treatment. C Immunoblots of total and phosphorylated (p) Akt in the uteri of vehicle (CT) and E3-treated mice (E3). D Heat maps showing the top 3000 differential CpG sites with altered methylation in E3-treated compared to control uterus. E Representative demethylated genes with altered expression from A and B. Error bars: mean + SEM. *, p < 0.05; **, p < 0.01; ***, p < 0.001 by Student t test, n = 8 mice/group. Programming of estrogen response: F, G Pregnant CD-1 mice were prenatally exposed to E3 (E3) or vehicle (CT). Six weeks after birth, the female offspring were ovariectomized; after 2 weeks they were transiently (6hrs) treated with either E2 or vehicle control as adults (designated CT-CT, CT-E2, E3-CT, and E3-E2, where the first term prior to the dash signifies prenatal exposure and the second term after the dash is the transient adult treatment). F, G Quantification of selected gene expression by quantitative RT-PCR. F The representative genes demonstrating an exaggerated estrogen response as adults after prenatal E3 exposure. G The genes where enhanced repression in response to E2 exposure was programmed by fetal E3 exposure. Error bars: mean + SEM. *, p < 0.05; **, p < 0.01; ***, p < 0.001 by Student t test, n = 12 mice/group. Uncropped blots available in Additional file 3: Fig. S7