Fig. 3. Dnmt2 RNA interference results in loss of embryonic DNA methylation. Embryos were injected with Dnmt2 dsRNA and then staged, fixed and double stained with antibodies against 5-methylcytosine (5mC, green) and DNA (red). The left panel shows an overlay of both the 5-methylcytosine and the DNA signal, the right panel shows the 5-methylcytosine signal only. Scale bars: 50 µm. (A) Injection of Dnmt2 dsRNA resulted in a complete loss of nuclear 5-methylcytosine signals. The signal for DNA was not affected. (B) Injection of CG11840 dsRNA revealed a nuclear staining similar to wild-type embryos. (C) The effect of RNA interference on embryonic development. The percentage of hatching first instar larvae (hatch rate) did not differ significantly between embryos injected with Dnmt2 dsRNA (black bar) and embryos injected with injection buffer only (white bar). Embryos injected with CG11840 dsRNA showed detectable embryonic lethality (grey bar). For each experiment, about 300 embryos were analyzed.