Journal: Stem Cell Research & Therapy
Article Title: Conditioned medium from bone marrow-derived mesenchymal stem cells inhibits vascular calcification through blockade of the BMP2–Smad1/5/8 signaling pathway
Figure Lengend Snippet: Noggin, BMP2 antagonist, suppresses VSMC calcification by inhibiting Smad1/5/8 signaling. VSMCs pretreated with Noggin (100 ng/ml) or vehicle for 2 h followed by incubation with 10 mM β-GP. a Alizarin Red staining (×100). b Quantification of Alizarin Red staining. c Calcium contents measured and normalized by protein content of cell lysates. mRNA expression of ( d ) BMP2, ( e ) Msx2, ( f ) Runx2, and ( g ) osteocalcin analyzed by rt-PCR. h Protein expression of BMP2 and phospho-Smad1/5/8 performed by western blot analysis. β-GP beta-glycerophosphate, DMEM Dulbecco’s Modified Eagle’s Medium, DMEM β-GP DMEM medium containing 10 mM β-GP, DMEM Noggin+β-GP DMEM medium containing 10 mM β-GP and 100 ng/ml Noggin, Msx2 msh homeobox 2, NC DMEM medium, OD optical density, Runx2 Runt-related transcription factor 2
Article Snippet: Beta-glycerophosphate (β-GP, G9422) and Alizarin Red S (A5533) were obtained from Sigma.
Techniques: Incubation, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Modification