Fig 3. Initial floxed Poldip2 mice carry a genomic duplication.

A: Copy number variation assays were performed by qPCR using two primer pairs and two TaqMan probes in each well to amplify both a reference gene and one short genomic segment at the indicated location (top). Bars are averages from quadruplicate representative samples ± confidence intervals calculated by CopyCaller software. Individual mouse ID numbers are indicated below each bar. WT is a wild type control and Em is an embryo. Apparent genotypes, obtained by conventional PCR as in Fig 2, are indicated at the bottom. The presence of 3 and 4 copies per genome is consistent with a duplication carried by heterozygotes (Het) and homozygotes, respectively. B: Increased expression of Poldip2 and neighboring genes in floxed mice. Poldip2 genotypes were determined by conventional PCR as in Fig 2, in addition to copy number variation assays, as in panel A, assuming that homozygotes (Hom) have 4 copies per genome. Total RNA was purified from duodenum. Expression of Poldip2 and adjacent genes Tmem199 and Tnfaip1, as well Nox4, a distant gene used as a control, was assessed by RT-qPCR and normalized to the wild type for each gene. Bars represent average ± SEM (n = 3–5 mice) **P< 0.01, ***P<0.001 vs. WT.