Sufficiency of the DR transgene for establishing and reprogramming imprinted DNA methylation during gametogenesis.

<p>Bisulfite analysis of DNA from sperm (A) or oocytes (B) of animals inheriting the DR transgene. Mice derived from male founder 2771 inherited the transgene from their mothers (2771/+) or fathers (+/2771); mice derived from female founder 3047 inherited the transgene from their mothers (3047/+) or fathers (+/3047). Transgene-bearing parent is indicated with a filled ♀ or ♂ symbol. Results are presented as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033024#pone-0033024-g003" target="_blank">Fig. 3</a>. Note that additional CpGs not in the endogenous DMD that were present on the transgene were also reprogrammed. C. DNA methylation in transgenic mice descended from a three generation pedigree from female founder 3047. Abbreviated pedigree (top) shows only transgene-carrying mice and the sex of the animal transmitting the transgene at each generation. Oocyte or tail DNAs from F3 transgenic mice were amplified before (−) or after (H) <i>Hha</i>I digestion. Tail DNA from wild type (+/+) was used as a negative control. The PCR was done using primers shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033024#pone-0033024-t002" target="_blank">Table 2</a> specific for the DMD of the transgene, which span five <i>Hha</i>I sites, or primers from actin, which span no <i>Hha</i>I sites. Gels (middle) or Q-PCR results (bottom) are shown with relative abundance of the signals from cut <i>vs.</i> uncut DMD amplifications indicated.