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. 2010 Jun 8;285(32):24783–24792. doi: 10.1074/jbc.M110.130286

FIGURE 6.

FIGURE 6.

Selected mutant RGMc proteins inhibit BMP-mediated gene expression. A, analysis of purified RGMc-Fc (R-Fc) fusion proteins or IgG1-Fc (Fc) after SDS-PAGE by Coomassie Blue staining (left panel), or by immunoblotting for IgG1-Fc (middle panel) or RGMc (right panel). RGMc proteins are as follows: 50, RGMc50; 40, RGMc40; G92V, RGMcG92V substitution mutant; G313V, RGMcG313V substitution mutant; 141X, RGMc141X truncation mutant. B, effects on gene expression: BMP2 or BMP6 (100 ng/ml) were preincubated for 3 h at 20 °C with 50-fold molar excess of IgG1-Fc, 4-fold molar excess of Noggin-Fc (Nog-Fc), 10-fold molar excess of R50-Fc or R40-Fc, or 4–20-fold molar excess of R-G92V-Fc, R-G313V-Fc, or R-141X-Fc, and then added to Hep3B cells for 4 h. Analysis of hepcidin, SMAD7, ID1, and S17 mRNA levels by RT-PCR is shown.