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. 2008 May 16;283(20):13874–13888. doi: 10.1074/jbc.M707820200

FIGURE 2.

FIGURE 2.

Velocity sedimentation analyses of BMP-7 complex formation. The ability of BMP-7 pd (expressed in bacteria) and separated BMP-7 gfd (expressed as a complex in 293 cells and separated as described (12)) to form a complex was tested using velocity sedimentation through sucrose gradients. A, control reference runs of separated BMP-7 pd (top left panel) and separated BMP-7 gfd (top right panel) show a peak around fraction 23 (immunoblotted with anti-BMP-7 pd antibody) and one around fractions 19–20 (immunoblotted with anti-BMP-7 gfd antibody). The control reference run for the BMP-7 complex (expressed in 293 cells) shows a peak around fraction 14 when detected with an anti-BMP-7 pd antibody (lower left panel) or with an anti-BMP-7 gfd antibody, indicating that BMP-7 pd and gfd migrate together as a complex through the gradient. B, BMP-7 complex formation after incubation of BMP-7 pd (expressed in bacteria) with separated BMP-7 gfd at a molar ratio of 2:1 (BMP-7 pd:BMP-7 gfd) was demonstrated by immunoblotting of fractions after velocity sedimentation using an anti-BMP-7 pd antibody (upper left panel) and an anti-BMP-7 gfd antibody (upper right panel). The BMP-7 pd and gfd signals were found in a similar position in the gradient as the native BMP-7 complex in the reference run (A), indicating the ability of the BMP-7 pd expressed in bacteria to form a complex with the BMP-7 gfd. The direction of sedimentation through the sucrose gradient is indicated under the fraction numbers (lower right panel).