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Figure 2. Grem2-BMP Binding Interface and Analysis
(A) Grem2-GDF5 complex with BMP-binding interfaces in Grem2 (labeled I through IV) highlighted in green, blue, red, and magenta, respectively.
(B–E) Grem2 and GDF5 are colored as in Figure 1. Zoomed-in view of the Grem2-GDF5 (B) interface I, (C) interface II, (D) interface III, and (E) interface IV, as described in Results.
(F) BMP-responsive luciferase reporter assay using a BMP-responsive osteoblast cell line and showing titration of various Grem2 mutants against 2 nM BMP2. Experiments were performed in triplicate. Error bars indicate mean ± SEM. WT, wild-type.
(G and H) Injection (stage 9) of wild-type and Grem2 mutants (0.5 μM, 1 μM, and 10 μM) in developing Xenopus embryos. Quantification (G) and representative images (H, top) of BMP-dependent axial development (stage 35). (H, bottom) In situ hybridization (stage 20) for mRNA expression of the BMP-target gene sizzled. Numbers in boxes indicate the concentration of the reagent used (top and bottom), as well as the number of embryos scored (bottom). |
