exogenous recombinant mouse bmp4  (R&D Systems)

Journal: Science Advances

Article Title: YAP regulates transcriptional programs for layer-specific periosteal expansion during fracture repair

doi: 10.1126/sciadv.adw0126

Figure Lengend Snippet: ( A ) Volcano plot of differentially accessible chromatin sites after 8 hours of YAP CA expression. Log 2 fold change > 2 indicates more accessible chromatin after YAP CA expression. ( B ) Motif analysis on differentially accessible chromatin loci. ( C ) Chromatin accessibility at locus proximate to Ctgf . ( D ) Chromatin immunoprecipitation of CTGF by YAP and TEAD in human cells. ( E ) Chromatin accessibility at locus proximate to Bmp4 . ( F ) Chromatin immunoprecipitation of BMP4 by YAP and TEAD in human cells. ( G ) Temporal dynamics of Yap and Bmp4 expression after 1, 4, 8, 12, 24, and 48 hours of doxycycline treatment. ( H ) Schematic for femoral osteotomies and periosteal cell isolation followed by MGH-CP1 treatment in vitro. ( I ) Bmp4 mRNA expression after 24 hours of treatment with 10 µm or 50 µm of MGH-CP1 in vitro. ( J ) Coimmunoprecipitation of TEAD with YAP in fracture-activated periosteal cells. IP, immunoprecipitation; IB, immunoblotting. ( K ) RNA in situ hybridization for Bmp4 in the periosteum of WT fl/fl and YAP/TAZcKO Osx mice. ( L ) Staining for Osx protein and DAPI in adjacent sections to sections in (K). Dotted lines indicate the periosteum. Scale bars, 100 μm. ** P < 0.01 and **** P < 0.0001.

Article Snippet: Recombinant mouse BMP4 (cat. no. 5020-BP, R&D Systems, Minneapolis, MN) was reconstituted in 0.9% sterile saline solution (cat. no. S5825, Teknova, Hollister, CA) at 12.5 μg/ml.

Techniques: Expressing, Chromatin Immunoprecipitation, Cell Isolation, In Vitro, Immunoprecipitation, Western Blot, RNA In Situ Hybridization, Staining

Journal: Science Advances

Article Title: YAP regulates transcriptional programs for layer-specific periosteal expansion during fracture repair

doi: 10.1126/sciadv.adw0126

Figure Lengend Snippet: ( A ) Timeline of in vivo recombinant mouse BMP4 (rmBMP4) delivery for 4 days after fracture in YAP/TAZ cKO Osx mice. ( B ) Immunofluorescence staining of Osx protein with DAPI and EdU. Dotted lines indicate the periosteum and the separation between the Osx + cambium and Osx − fibrous layers. ( C ) Average periosteal thickness. ( D ) Percentage of proliferative EdU + cells in the periosteum. ( E ) Quantification of cambium and fibrous layer thickness. ( F ) Percentage of proliferative EdU + cells in each layer. Scale bars, 100 μm. * P < 0.05, ** P < 0.01.

Article Snippet: Recombinant mouse BMP4 (cat. no. 5020-BP, R&D Systems, Minneapolis, MN) was reconstituted in 0.9% sterile saline solution (cat. no. S5825, Teknova, Hollister, CA) at 12.5 μg/ml.

Techniques: In Vivo, Recombinant, Immunofluorescence, Staining

Journal: Science Advances

Article Title: YAP regulates transcriptional programs for layer-specific periosteal expansion during fracture repair

doi: 10.1126/sciadv.adw0126

Figure Lengend Snippet: ( A ) ALP activity in the periosteum of YAP/TAZ cKO Osx mice treated with saline or BMP4 at 4 DPF. ( B and C ) Quantification of ALP intensity (B) and ALP + area fraction (C). ( D ) Blood vessels marked by endomucin (Emcn) staining. ( E ) Frequency distribution of blood vessel loop size. ( F ) SHG imaging of collagen matrix content and organization. ( G ) Quantification of SHG signal. Scale bars, 100 μm. * P < 0.05.

Article Snippet: Recombinant mouse BMP4 (cat. no. 5020-BP, R&D Systems, Minneapolis, MN) was reconstituted in 0.9% sterile saline solution (cat. no. S5825, Teknova, Hollister, CA) at 12.5 μg/ml.

Techniques: Activity Assay, Saline, Staining, Imaging

Journal: Science Advances

Article Title: YAP regulates transcriptional programs for layer-specific periosteal expansion during fracture repair

doi: 10.1126/sciadv.adw0126

Figure Lengend Snippet: ( A ) Timeline of in vivo recombinant mouse BMP4 delivery for 4 days after fracture in WT YAP/TAZ fl/fl mice. ( B ) Immunofluorescence staining of Osx protein with DAPI and EdU. Dotted lines indicate the periosteum and the separation between the Osx + cambium and Osx − fibrous layers. ( C ) Average periosteal thickness. ( D ) Percentage of proliferative EdU + cells in the periosteum. ( E ) Quantification of cambium and fibrous layer thickness. ( F ) Percentage of proliferative EdU + cells in each layer. Scale bars, 100 μm.

Article Snippet: Recombinant mouse BMP4 (cat. no. 5020-BP, R&D Systems, Minneapolis, MN) was reconstituted in 0.9% sterile saline solution (cat. no. S5825, Teknova, Hollister, CA) at 12.5 μg/ml.

Techniques: In Vivo, Recombinant, Immunofluorescence, Staining

Journal: Science Advances

Article Title: YAP regulates transcriptional programs for layer-specific periosteal expansion during fracture repair

doi: 10.1126/sciadv.adw0126

Figure Lengend Snippet: ( A ) ALP activity in the periosteum of YAP fl/fl TAZ fl/fl mice treated with saline or BMP4 at 4 DPF. ( B and C ) Quantification of ALP intensity (B) and ALP + area fraction (C). ( D ) Blood vessels marked by endomucin staining. ( E ) Frequency distribution of blood vessel loop size. K-S, Kolmogorov-Smirnov. ( F ) SHG imaging of collagen matrix content and organization. ( G ) Quantification of SHG signal. Scale bars, 100 μm. * P < 0.05, ** P < 0.01.

Article Snippet: Recombinant mouse BMP4 (cat. no. 5020-BP, R&D Systems, Minneapolis, MN) was reconstituted in 0.9% sterile saline solution (cat. no. S5825, Teknova, Hollister, CA) at 12.5 μg/ml.

Techniques: Activity Assay, Saline, Staining, Imaging