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DmelNV VP1 interacts with Dmel AGO2 in S2 cells.

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posted on 17.07.2014, 03:17 authored by Joël T. van Mierlo, Gijs J. Overheul, Benjamin Obadia, Koen W. R. van Cleef, Claire L. Webster, Maria-Carla Saleh, Darren J. Obbard, Ronald P. van Rij

(A) Western blot (WB) analysis of V5 immunoprecipitation on lysates from S2 cells transfected with a FLAG-AGO2 expression plasmid and either V5-tagged DmelNV VP1 (V5-VP1) or V5-control plasmid (Vector). The epitope-tagged proteins were detected in the input, supernatant after immunoprecipitation (Sup), and the immunoprecipitate (V5-IP) with the indicated antibodies. (B) FLAG immunoprecipitation of lysates from S2 cells transfected with V5-tagged DmelNV VP1 (V5-VP1) and either FLAG-AGO2 or FLAG-control plasmids (Vector), followed by western blot analysis with the indicated antibodies. (C) V5 immunoprecipitation of lysates from S2 cells transfected with V5-tagged DmelNV VP1 (+) or V5-control (−) plasmids. After SDS-PAGE, endogenous AGO2 or DmelNV VP1 proteins were detected by western blot using anti-AGO2 (α-AGO2) and anti-V5 (α-V5) antibody, respectively. Asterisk (*) indicates a non-specific background band; triangle indicates AGO2. The DmelNV VP1ΔN351 construct was used in these experiments.

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