Paternally Expressed Peg3 Controls Maternally Expressed Zim1 as a Trans Factor
Figure 5
Restoring the protein levels of PEG3 down-regulates Zim1.
(A) Genomic structure of the mutant allele of Peg3 and FLP-mediated recombination scheme to restore the expression of Peg3. The inserted cassette is flanked by two FRT sites, thus can be removed by Flippase (FLP). (B) Three pools of KO MEF cells were transfected with the following constructs: no vector as a mock control (lane 1), a Green Fluorescent Protein (GFP) expression vector as a negative control (lane 2), and a FLP expression vector (lane 3). The total RNA isolated from these cells were analyzed with RT-PCR to measure the expression levels of β-actin, Zim1, and Peg3. The bottom panel shows genotyping results confirming FLP-mediated recombination of the mutant allele (Rev KO) and endogenous allele (WT) of Peg3. (C) The observed down-regulation of Zim1 was further analyzed using qRT-PCR.