Figure 4

Dnmt3a , Dnmt3b and Dnmt3L expression in DNMT3L deficient 15 dpp oocytes and in DNMT1o deficient 25 dpp GV oocytes. a) QRT-PCR was used to analyze the expression of the de novo DNMT enzymes in Dnmt3L heterozygous (dark grey bars) and homozygous (cross hatch bars) oocytes at 15 dpp. Results for Dnmt3a and Dnmt3b are shown in a) while relative expression of Dnmt3L is illustrated in b). Dnmt3a and Dnmt3b transcripts are up-regulated in DNMT3L depleted growing oocytes. The differences observed for Dnmt3b and Dnmt3L were statistically significant (p < 0.01). c) QRT-PCR was used to analyze the expression of the DNMT enzymes in Dnmt1o heterozygous (dark grey bars) and homozygous (cross hatch bars) 25 dpp GV stage oocytes. While the relative expression of Dnmt3a and Dnmt3b was up-regulated in DNMT1o depleted GV oocytes, Dnmt3L expression remained unchanged. Samples were analyzed in triplicate and relative expression values obtained were normalized to the level of rabbit α-globin expression for each sample and were calibrated to the expression in heterozygous oocytes for Dnmt3a, Dnmt3b and Dnmt3L. For Dnmt3L analysis, expression was calibrated to the expression in homozygous oocytes. Results for one experiment are presented as mean ± SD.