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. 2011 Sep 20;286(45):38939–38947. doi: 10.1074/jbc.M111.291377

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — MAM7, fibronectin, and phosphatidic acid form a tripartite complex. Shown are pulldown assays using GST-MAM7 or -MAM1, fibronectin, and liposomes (with or without PA). GST-MAM proteins were incubated with fibronectin followed by liposomes (A and C) or liposomes followed by fibronectin (B and D). Bound proteins were analyzed by SDS-PAGE and Coomassie staining (A and B). Liposomes in flow-through and eluate fractions were quantitated using a molybdophosphoric acid assay and expressed as fractions of loaded lipid (C and D). Alexa Fluor 488-labeled MAM7 was tested for binding to Fn-coated plates in the presence of increasing concentrations of liposomes prepared from PC (●) or a mixture containing 20:80 mol % PC:PA (○) (E). Samples incubated with the highest concentration (conc.) of liposomes (arrow) were analyzed for bound liposomes using the molybdophosphoric acid assay and compared with plate assays carried out with MBP control (F).