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FIG. 3. DRAGON interacts directly with BMP2 and BMP4. A, DRAGON expressed in COS cells binds to BMP2 but not TGF . COS-1 cells were transiently transfected with DRAGON (DR), HA-tagged ALK6 (A6/HA), or T RII (RII/HA). Cells were affinity-labeled with [125I]-BMP2 (left panel) or [125I]-TGF (right panel) and lysates subjected to immunoprecipitation (IP) or immunoblotting with the indicated antibodies. 125I-Labeled ligand binding to receptors was detected by phosphorimaging. B, DRAGON-Fc binds directly to BMP2. [125I]-BMP2 was incubated with increasing amounts (10–50 ng) of purified DRAGON-Fc then added to protein A-coated plates, and bound radioactivity was measured (left graph). 20 ng of DRAGON-Fc were incubated with [125I]-BMP2 in the presence of increasing amounts of competing unlabeled BMP2 (0–42 nM) (right graph). C, DRAGON-Fc binds directly to BMP2 and BMP4 but not BMP7 or TGF ligands. DRAGON-Fc was incubated with [125I]-BMP2 in the presence of unlabeled BMP2, BMP4, BMP7, Activin A (ActA), TGF 1, TGF 2, and TGF 3, and [125I]-BMP2 binding was measured. D, DRAGON-Fc reduces BMP2 mediated signaling. LLC-PK1 cells were transiently transfected with BRE-Luc construct and treated with BMP2 (150 ng/ml) and DRAGON-Fc (60 and 300 ng/ml). E, DRAGON-Fc (60 ng/ml) reduces BMP2 but not BMP7- or TGF 1 (1 nM each)-mediated signaling. LLC-PK1 cells were transiently transfected with BRE-Luc and treated with BMP2 or BMP7 or transiently transfected with 3TP-Lux construct and treated with TGF 1 in the absence or presence of DRAGON-Fc. Image published in: Samad TA et al. (2005) Copyright © 2005. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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