Figure 4.

Mutational effects on transcriptional levels. A series of qRT-PCR were performed to test potential effects of the mutation on the transcriptional levels of the imprinted genes within the Peg3 domain. This series of analyses used the total RNA isolated from the brain of the one-day-old pups carrying the mutant allele paternally (A,C) and maternally (B). The expression levels of each gene were first normalized with an internal control (β-actin), and the normalized values were further compared between the wild-types and the heterozygotes. The relative levels are presented in a graph with standard errors (S.E.). A similar series of expression analyses were also conducted using a set of 14.5-dpc placenta (D). Statistical significance by Student’s t-test was indicated in the following manner: * < 0.05, ** < 0.01, *** < 0.001. The series of analyses were repeated with two independent sets of biological replicates.