rabbit polyclonal anti phospho smad1 5 8  (Cell Signaling Technology Inc)

Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Palovarotene inhibits osteochondroma formation in a mouse model of multiple hereditary exostoses †

doi: 10.1002/jbmr.3341

Figure Lengend Snippet: Fsp1Cre;R26tdTomato;Ext1flox/flox (CKO) mice were treated by daily oral gavage of 1.76 mg/kg PVO or vehicle for 5 days starting at P21. At P26, these mice as well as vehicle-treated Fsp1Cre;R26tdTomato (WT) mice were sacrificed, and frozen sections of the 10th ribs were stained with anti-pSmad1/5/8 antibody (green). Fsp1Cre-mediated recombination was monitored by tdTomato (red). (A) pSmad1/5/8 expression in the perichondrium. (B) pSmad1/5/8 expression in the periosteum. Note that only low levels of pSmad1/5/8 are seen in the perichondrium and periosteum of vehicle-treated WT mice (WT/Vehicle), whereas Smad1/5/8 is strongly phosphorylated in vehicle-treated CKO mice (CKO/Vehicle). In PVO-treated CKO mice, Smad1/5/8 phosphorylation is attenuated to the WT level (CKO/PVO). Data shown are representative images. Each analysis was performed on at least 3 animals per genotype. Scale bars, 50 μm (A and B). (C) Ext1-deficient (Ext1−/−) and wild-type (WT) PDPCs in monolayer cultures were pretreated with 1 μM PVO in DMSO or DMSO only for 48 h. Then, cells were incubated with 10 ng/ml BMP2 for 30 min and lysed. Cell lysates were immunoblotted with antibodies to pSmad1/5/8, total Smad1/5/8, and α-tubulin. This experiment was performed three times with similar results.

Article Snippet: Western blotting was performed with rabbit polyclonal anti-phospho-Smad1/5/8 antibody (described above), rabbit polyclonal anti-Smad1/5/8 (Cat. No. sc-6031-R) from Santa Cruz Biotechnology (Dallas, TX), and mouse monoclonal anti-α-tubulin antibody (Cat. No. T6074) from Sigma-Aldrich (St. Louis, MO).

Techniques: Staining, Expressing, Incubation