Journal: EMBO Molecular Medicine

Article Title: Supraphysiological levels of GDF 11 induce striated muscle atrophy

doi: 10.15252/emmm.201607231

Figure Lengend Snippet: Relative p‐SMAD2/3 (left) and p‐SMAD1/5/8 (right) response, evaluated by AlphaLISA signal, of HEK293T cells after 1‐h exposure to recombinant myostatin (Mstn), activin A, GDF11, TGFβ, and BMP2. Relative p‐SMAD2/3 response of C2C12 myoblasts (left) and myotubes (right) after 1‐h exposure to Mstn, activin A, GDF11, and TGFβ. Phosphorylation of SMAD2 and SMAD3 in differentiated C2C12 myotubes following stimulation with recombinant Mstn or GDF11 for 30 and 60 min, as detected by immunoblotting. Equal loading is verified by Ponceau Red staining. EC 50 values (in nM) for p‐SMAD2/3 and p‐SMAD1/5/8 responses of HEK293T, C2C12 myoblasts, and C2C12 myotubes to the above listed ligands, as well as p‐SMAD1/5/8 response to BMP4, BMP6, and BMP7. Data information: Values are displayed as mean ± SEM; n = 4 for all data points.

Article Snippet: To assess the phosphorylation of SMAD in response to TGFβ superfamily members in undifferentiated cells, C2C12 myoblasts (ATTC CRL‐1772; passage 14) and HEK293T cells (ATCC CRL‐11268; passage 13) were plated on 96‐well tissue culture‐treated plates (Greiner‐Bio‐One 655098) in growth medium [high glucose Dulbecco's modified Eagle's medium (DMEM) + 10% fetal bovine serum (FBS) + 1% penicillin streptomycin (PS)], grown to confluency, and were treated with various doses of the following recombinant ligands for 1 h ( n = 4): Mstn (R&D Systems #788‐G8), GDF11 (R&D Systems # 1958‐GD), TGFβ (R&D Systems #240‐B), activin A (R&D Systems # 338‐AC), BMP‐2 (R&D Systems # 355‐BM), BMP‐4 (R&D Systems # 314‐BP), BMP‐6 (R&D Systems # 507‐BP), or BMP‐7 (R&D Systems # 354‐BP).

Techniques: Recombinant, Phospho-proteomics, Western Blot, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Berberine and aspirin prevent traumatic heterotopic ossification by inhibition of BMP signalling pathway and osteogenic differentiation

doi: 10.1111/jcmm.17919

Figure Lengend Snippet: The effect of berberine and aspirin on the BMP signalling pathway. C3H10T1/2 cells were treated for 24 h with only berberine or aspirin with different doses (A, B), or followed by BMP6 (20 ng/mL) for 30 min (C, D). Western blot was performed with indicated antibodies. Each experiment was repeated three times independently and the representative blots were shown in the figure.

Article Snippet: Recombinant mouse BMP‐6 protein was obtained from R&D System.

Techniques: Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Berberine and aspirin prevent traumatic heterotopic ossification by inhibition of BMP signalling pathway and osteogenic differentiation

doi: 10.1111/jcmm.17919

Figure Lengend Snippet: The combination effects of berberine and aspirin on the BMP signalling pathway. (A, B). C3H10T1/2 cells were treated with berberine and aspirin alone or in combination for 24 h. (C, D). The cells were treated with berberine and aspirin alone or in combination for 24 h, followed by BMP6 (20 ng/mL) for 30 min. Western blot was performed with indicated antibodies. Each experiment was repeated three times independently and the representative blots were shown in the figure.

Article Snippet: Recombinant mouse BMP‐6 protein was obtained from R&D System.

Techniques: Western Blot