FIGURE 5:

miR-322/503–modulated expression of Smurf2 regulates the degradation of phosphorylated Smad2. (A) Levels of phosphorylated Smad2 (p-Smad2) and total Smad2 (T-Smad2) in cells treated with TGF-β (1 ng/ml) for different times. The levels of p-Smad2 and T-Smad2 were measured by Western blotting analysis using anti–p-Smad2 or anti-Smad2 antibody. (B) Changes in the levels of p-Smad2 and T-Smad2 in cells overexpressing GFP-tagged Smurf2 after treatment with TGF-β. After cells were transfected with the Smurf2 expression vector or control vector for 24 h, they were exposed to TGF-β for 0.5 or 1 h. (C) Levels of p-Smad2 and T-Smad2 48 h after cotransfecting cells with anti–miR-322 and control siRNA (C-siRNA) or siRNA targeting Smurf2 mRNA (siSmurf2). (D) Changes in the levels of p-Smad2 and T-Smad2 in miR-322–silenced cells exposed to TGF-β. Cells were initially transfected with anti–miR-322 for 48 h and then exposed to TGF-β for 0.5 or 1 h. (E) Levels of p-Smad2 and T-Smad2 48 h after cotransfecting cells with anti–miR-503 and siSmurf2 for 48 h. (F) Changes in the levels of p-Smad2 and T-Smad2 in miR-503–silenced cells exposed to TGF-β for different times.