Figure 1
From: Blockade of Smad signaling by 3′-deoxyadenosine: a mechanism for its anti-fibrotic potential
Suppression of TGF-β and BMP-4 signaling by 3′-deoxyadenosine (3′-DA). (a) NRK-52E cells were treated with 5 ng/ml TGF-β1 or 50 ng/ml BMP-4 in the absence (−) or presence (+) of 5 μg/ml 3′-DA for 24 h, and expression of type I collagen (Col I) and type IV collagen (Col IV) was evaluated by northern blot analysis. The level of GAPDH mRNA serves as a loading control. Quantitative assessment of Col I mRNA level normalized by the level of GAPDH is shown in the graph. (b) Cells were exposed to TGF-β1 or BMP-4 in the presence of serial concentrations of 3′-DA for 24 h and subjected to northern blot analysis. Quantitative assessment is shown in graphs. (c–e) Cells were transfected with p(CAGA)12-MLP-Luc (c), pBRE-Luc (d) or pSV40-Luc (e), treated with indicated reagents for 24 h and subjected to chemiluminescent assay. (f, g) Cells were transfected with p(CAGA)12-MLP-Luc (f) or pBRE-Luc (g), treated with adenosine (ADO; 100 μM), ADP (100 μM) or ATP (100 μM) in the presence of TGF-β (f) or BMP-4 (g) for 24 h and subjected to chemiluminescent assay. Data are expressed as means±s.e., and asterisks indicate statistically significant differences (P<0.05). NS, not statistically significant; RLU, relative light unit.
