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Initial and final structures of Noggin-BMP-2 and Noggin-BMP-9 after 100 ns MD simulation. Identification of the binding effect of BMP-2 and BMP-9 to BMPR2 <t>in</t> <t>MC3T3-E1</t> Cells by Noggin using Immunoprecipitation. ( A ) Initial structures of Noggin-BMP-2 and ( B ) Initial structures of Noggin-BMP-9 ( C ) Aligned structures of initial (White color) and final structures of Noggin-BMP-2 (Coral/Blue) ( D )Aligned structures of initial (White) and final structures of Noggin-BMP-9 (Green/Ultraviolet) ( E ) RMSD of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). ( F ) RMSF (Root Mean Squared Fluctuation) of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). RMSD and RMSF values ​​represent the mean and standard deviation of three independent simulations. ( G ) Western blot analysis using total cell lysates. ( H ) Immunoprecipitation (IP) of BMPR2 followed by immunoblotting (IB) to detect BMP-2 and BMP-9 binding.
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Initial and final structures of Noggin-BMP-2 and Noggin-BMP-9 after 100 ns MD simulation. Identification of the binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 Cells by Noggin using Immunoprecipitation. ( A ) Initial structures of Noggin-BMP-2 and ( B ) Initial structures of Noggin-BMP-9 ( C ) Aligned structures of initial (White color) and final structures of Noggin-BMP-2 (Coral/Blue) ( D )Aligned structures of initial (White) and final structures of Noggin-BMP-9 (Green/Ultraviolet) ( E ) RMSD of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). ( F ) RMSF (Root Mean Squared Fluctuation) of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). RMSD and RMSF values ​​represent the mean and standard deviation of three independent simulations. ( G ) Western blot analysis using total cell lysates. ( H ) Immunoprecipitation (IP) of BMPR2 followed by immunoblotting (IB) to detect BMP-2 and BMP-9 binding.

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Initial and final structures of Noggin-BMP-2 and Noggin-BMP-9 after 100 ns MD simulation. Identification of the binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 Cells by Noggin using Immunoprecipitation. ( A ) Initial structures of Noggin-BMP-2 and ( B ) Initial structures of Noggin-BMP-9 ( C ) Aligned structures of initial (White color) and final structures of Noggin-BMP-2 (Coral/Blue) ( D )Aligned structures of initial (White) and final structures of Noggin-BMP-9 (Green/Ultraviolet) ( E ) RMSD of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). ( F ) RMSF (Root Mean Squared Fluctuation) of Noggin-BMP-2 and Noggin-BMP-9 complexes during the all-atom MD simulation (100 ns). RMSD and RMSF values ​​represent the mean and standard deviation of three independent simulations. ( G ) Western blot analysis using total cell lysates. ( H ) Immunoprecipitation (IP) of BMPR2 followed by immunoblotting (IB) to detect BMP-2 and BMP-9 binding.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Immunoprecipitation, Standard Deviation, Western Blot

Identification of binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 cells by Noggin. ( A ) BMP-2–treated group, ( B ) BMP-2 + Noggin–treated group, ( C ) BMP-9–treated group, ( D ) BMP-9 + Noggin–treated group. DAPI (blue), anti-BMPR2 (red), and anti-BMP-2 or anti-BMP-9 (green). Merged images show co-localization (yellow). Scale bar: 10 μm; magnification: ×600. ( E ) Western blot of SMAD1/5/9 phosphorylation under the indicated treatments. Top, p-SMAD1/5/9; middle, total SMAD1/5/9; bottom, GAPDH. ( F) Alkaline phosphatase (ALP) activity measured 60 min after BMP addition using a p-nitrophenyl phosphate colorimetric assay (405 nm). Bars indicate mean ± SD ( n ≥ 3). p < 0.0001 vs. NT for all treated groups; asterisks denote significance relative to NT.

Journal: Scientific Reports

Article Title: Understanding of the different roles of Noggin in the Noggin-BMP-2 and Noggin-BMP-9 dimer complexes at the molecular level

doi: 10.1038/s41598-025-33735-8

Figure Lengend Snippet: Identification of binding effect of BMP-2 and BMP-9 to BMPR2 in MC3T3-E1 cells by Noggin. ( A ) BMP-2–treated group, ( B ) BMP-2 + Noggin–treated group, ( C ) BMP-9–treated group, ( D ) BMP-9 + Noggin–treated group. DAPI (blue), anti-BMPR2 (red), and anti-BMP-2 or anti-BMP-9 (green). Merged images show co-localization (yellow). Scale bar: 10 μm; magnification: ×600. ( E ) Western blot of SMAD1/5/9 phosphorylation under the indicated treatments. Top, p-SMAD1/5/9; middle, total SMAD1/5/9; bottom, GAPDH. ( F) Alkaline phosphatase (ALP) activity measured 60 min after BMP addition using a p-nitrophenyl phosphate colorimetric assay (405 nm). Bars indicate mean ± SD ( n ≥ 3). p < 0.0001 vs. NT for all treated groups; asterisks denote significance relative to NT.

Article Snippet: The mouse pre-osteoblastic cell line MC3T3-E1 was purchased from the American Type Culture Collection (ATCC, CRL-2593, Manassas, VA, USA).

Techniques: Binding Assay, Western Blot, Phospho-proteomics, Activity Assay, Colorimetric Assay