Novel Drosophila Viruses Encode Host-Specific Suppressors of RNAi
(A) V5 Immunoprecipitation (V5-IP) of lysates from S2 cells transfected with FLAG-tagged Dmel AGO2 expression plasmid and either V5-tagged DmelNV VP1, DimmNV VP1, or V5-control plasmids (−). Input, supernatant after immunoprecipitation (Sup), and the immunoprecipitate (V5-IP) were analyzed by western blot (WB) using anti-V5 (α-V5) or anti-FLAG (α-FLAG) antibodies. (B) V5 immunoprecipitation of S2 cells transfected with plasmids encoding V5-tagged DmelNV VP1, DimmNV VP1, or V5-control vector (−). Input, sup, and IP fractions were analyzed by western blot using antibodies for endogenous AGO2 (α-Dmel AGO2) and V5 (α-V5). (C) V5 immunoprecipitation on lysates from S2 cells co-transfected with plasmids encoding FLAG-tagged Dimm AGO2 and either V5-tagged DmelNV VP1, DimmNV VP1, or V5-control vector (−). VP1 and Dimm AGO2 proteins were detected on western blot using anti-V5 (α-V5) and anti-FLAG (α-FLAG) antibodies, respectively. Asterisks (*) indicate a non-specific background band; triangles indicate AGO2. For these experiments the corresponding DmelNV VP1ΔN284 and DimmNV VP1ΔN295 constructs were used (Figure S1).