Figure 5

Protein and transcript variant analysis of Dnmt3a and Dnmt3b in DNMT3L wild type and depleted 15 dpp oocytes. Western Blot analysis of a) DNMT3A and c) DNMT3B was carried out on growing oocytes isolated from Dnmt3L wild-type and homozygous 15 dpp females (280 oocytes/lane). For comparison and isoform identification, the panels on the right labelled 'B' show blotting results for type B spermatogonia (for the loading control see Additional file 4). As suggested by the RNA expression analysis in (b), both DNMT3a and DNMT3a2 are present in growing oocytes and the ratio of these transcripts does not change significantly in Dnmt3L homozygous oocytes. DNMT3b2 is the more abundant isoform in wild-type Dnmt3L oocytes and appears to be upregulated in mutant oocytes as also suggested by the RNA expression data in Figure 4a. b) Relative expression of Dnmt3a and Dnmt3a2 was determined using transcript specific primers. Expression was normalized to 18S. As previously mentioned, both transcripts are expressed in growing oocytes.