Figure 7

Imprinted gene expression in DelTel7/IC2KO placentae. The expression of three IC2-regulated genes was analyzed in wild type (+/+) and rescued (DelTel7/IC2KO) placentae on placental section (top) and by qRT-PCR on total placental RNA (bottom). A) Phlda2 expression analyzed by IHC on paraffin sections of wild type (n = 4) and mutant (n = 4) placentae at E9.5. The qRT-PCR was performed on E9.5 placental RNA; DelTel7/IC2KO and +/+. B) Ascl2 expression analyzed by ISH on frozen placental sections at E14.5; n = 4 for DelTel7/IC2KO and wild type. Ascl2 expression by qRT-PCR at E14.5 on mutant placentae compared with +/+. C) Cdkn1c expression in DelTel7IC2KO placentae (n = 9) at E14.5 compared with +/+ (n = 9). Cdkn1c expression by qRT-PCR at E14.5 on mutant placentae compared with +/+. Sense probes not shown. GCs = glycogen cells; lab = labyrinth; sp = spongiotrophoblast; Gi = trophoblast giant cells. The blue and brown stains show gene and protein expression, respectively. All scale bars: 0.25 mm. All qRT-PCRs at E9.5 were performed on five DelTel7/IC2KO placentae and five wild type placentae with three technical replicates per individual placentae. All qRT-PCRs at E14.5 were performed on three DelTel7/IC2KO placentae and three wild-type placentae with three technical replicates per individual placentae. All expression results by qRT-PCR are shown normalized to the Ppia reaction and results are shown ± SD. First, the average from each placenta was calculated from technical replicates from the same cDNA. Then the SD was calculated from the average of biological replicates per genotype. For the three genes tested the differences between wild type and mutant placentae are not statistically significant (p > 0.7). D) qRT-PCR of the other imprinted genes in the IC2 domain, Tssc4, Kcnq1 and Cd81, in E9.5 DelTel7/IC2KO placentae reveals no overall difference from wild type (p > 0.2). qRT-PCR was performed as described above on five placentae of each genotype.