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. 2011 Jun 23;7(6):e1002092. doi: 10.1371/journal.ppat.1002092

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Derré et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Schematic representation of the incDEFG operon and the IncD protein. (B) Lysates from HEK293 cells co-expressing 3xFLAG-IncD and the indicated GFP-CERT fusion proteins were immunoprecipitated with anti-FLAG M2 beads. A portion of the cell lysate (Left Panel, 2% Total) and the immunoprecipitated proteins (Right Panel, IP) were separated by SDS-PAGE and analyzed by immunoblot with antibodies against GFP (Top Panels) and FLAG (Bottom Panels). (C) Lysates from HEK293 cells co-expressing 3xFLAG-IncD, 3xFLAG-IncE or 3xFLAG-IncF and GFP-CERT fusion proteins were immunoprecipitated with anti-FLAG M2 beads. A portion of the cell lysate (Right Panel, 2% Total) and the immunoprecipitated proteins (Left Panel, IP) were separated by SDS-PAGE and analyzed by immunoblot with antibodies against GFP (Top Panels) and FLAG (Bottom Panels). (D) Lysates from HEK293 cells co-expressing 3xFLAG-IncD and the indicated GFP or YFP fusion proteins were immunoprecipitated with anti-FLAG M2 beads. A portion of the cell lysate (Right Panel, 2% Total) and the immunoprecipitated proteins (Left Panel, IP) were separated by SDS-PAGE and analyzed by immunoblot with antibodies against GFP (Top Panels) and FLAG (Bottom Panels). (E) The indicated GST fusion proteins, immobilized onto glutathione sepharose were incubated in the presence of the indicated purified MBP fusion proteins. The protein complexes bound to the resin were separated by SDS-PAGE and analyzed by immunoblot with antibodies against MBP (Top Panels) and GST (Bottom Panels). The inputs for each purified protein are shown in the right panels (Input).