<i>Dlk1</i> is expressed but not imprinted from the 127H5 transgene.

<p>(A) Schematic of the <i>Dlk1</i>-<i>Meg3</i> BAC clones used to generate transgenic mice; the <i>28G5</i> transgene was described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036483#pone.0036483-Yevtodiyenko2" target="_blank">[46]</a>. The 127H5 BAC was linearized at a unique <i>Cla</i>I site. (B, C) Representative Northern blots for <i>Dlk1</i> mRNA in midgestation wild type (WT) and heterozygous transgenic (Tg) embryo (B) and placenta (C), after paternal (<i>127H5<sup>Pat</sup></i>) and maternal (<i>127H5<sup>Mat</sup></i>) inheritance. The mouse <i>β-actin</i> gene was used as a loading control. (D) Quantitative Northern data for blots shown in B & C; expression is normalized to <i>β-actin</i>. Gray bars represent wild type samples and black bars represent <i>127H5</i> transgenic samples upon paternal (<i>127H5<sup>Pat</sup></i>) or maternal (<i>127H5<sup>Mat</sup></i>) transmission in crosses to Cg12. (E) Direct sequencing assay for <i>Dlk1</i> imprinting in wild type (WT) and heterozygous transgenic (Tg) F<sub>1</sub> embryos after paternal (<i>127H5<sup>Pat</sup></i>) and maternal (<i>127H5<sup>Mat</sup></i>) inheritance. D indicates wild type animals carrying only the <i>M. domesticus</i> allele, while D×C or C×D indicates offspring of crosses to the Cg12 line carrying a <i>M. castaneus</i> allele, with the female genotype listed first. (F) Quantitative Northern blot analysis for <i>Dlk1</i> mRNA in 3–4 week old <i>127H5</i> tissues. Expression is normalized to <i>β-actin</i>, and each bar represents 8–10 animals. Gray bars represent wild type samples and the black bars represent <i>127H5</i> transgenic samples upon paternal (Pat) or maternal (Mat) transmission. In all figures asterisks indicate p≤0.05.</p>