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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 3. — A minimum of five mce domains is required for stable binding of fibronectin to MAM. Pulldown experiments were performed using intact fibronectin and GST-MAM1, -MAM2, -MAM4, -MAM5, -MAM6, or -MAM7. Load and eluate fractions were analyzed by SDS-PAGE and visualized by Coomassie staining (A). Binding of labeled MBP-MAM1, -MAM2, -MAM6, and -MAM7 to Fn-coated plates was determined using fluorescence saturation binding assays (B). Shown are pulldown assays with GST-tagged individual mce domains (mce1 to mce7) and fibronectin. Load and eluate fractions were analyzed by SDS-PAGE and visualized by Coomassie staining (C). Shown are pulldown assays with GST-tagged mce1–5, -2–6, and -3–7 proteins and fibronectin (D), or GST-tagged (mce1)₃, (mce1)₅, and (mce1)₇ concatemers and fibronectin (E). conc., concentration.