Dnmt2 mutants show delayed immune responses and Dnmt2–EGFP relocalizes and interacts with DCV RNA during infection. Dnmt2 mutants show delayed immune responses.

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Dnmt2 mutants show delayed immune responses and Dnmt2–EGFP relocalizes and interacts with DCV RNA during infection. Dnmt2 mutants show delayed immune responses and Dnmt2–EGFP relocalizes and interacts with DCV RNA during infection. (A) Q‐PCR analysis for immune response genes in wild‐type (D2+/+), Dnmt2 mutant (D2−/−) and transgenic rescue (D2‐TG) recipient flies after feeding with donors and incubation periods as described in Fig 3A. (B) Northern blotting for Metchnikowin RNA in wild‐type (D2+/+) and Dnmt2 mutant (D2−/−) recipients after feeding with donors as described in Fig 3A and an incubation period of 1 day. Rp49 was probed as loading control. (C) Immunofluorescence images of fixed Drosophila hindgut tissue from D2‐TG recipients after feeding with yeast alone (yeast) or Dnmt2 mutant (D2−/−) donors followed by an incubation period of 1 day. The border between lumen and gut cells is shown. Dnmt2–EGFP fusion protein was visualized using antibodies against EGFP (white in upper panels, green in lower panels). DNA is shown in red. (D) Schematic illustration of the DCV RNA genome with three primer sets used for the analysis of RIPs. Agarose gels show the results of RIP from control (Ubq::EGFP) and D2‐TG (D2‐EGFP) recipients after continuous feeding (72 h) with Dnmt2 mutant donors. Both samples contained tRNA‐AspGTC (grey arrowhead) and DCV in the input fraction. After RIP, only Dnmt2–EGFP complexes contained tRNA‐AspGTC (grey arrowhead) and DCV. RNA expression was set to 1 in non‐infected recipients (ctrl) and normalized to rp49 mRNA in individual experiments. Error bars represent s.d.'s from three biological replicates. DCV, Drosophila C virus; Q‐PCR, quantitative PCR; RIP, RNA immunoprecipitation; rel., relative. Zeljko Durdevic et al. EMBO Rep. 2013;14:269-275 © as stated in the article, figure or figure legend