Figure 2
From: MiR-21/Smad 7 signaling determines TGF-β1-induced CAF formation
Smad 7 protects Smad 2/3 from activation through physical binding.
(A). Smad 7 regulates Smad 2/3 activation through physical binding. The top left panel shows Western blots of Smad 7 after Smad 2 or Smad 3 immunoprecipitation (IP), the top middle panel is the Western blots for the reciprocal IP and loading control (the full-length blots can be found in the Supplementary data Figure S1); the top right panel shows the effects of Smad 7 knock down on Smad 2/3 phosphorylation after TGF-β1 stimulation, the bottom left panel shows the phosphorylation kinetics of Smad 2 and Smad 3 after TGF-β1 stimulation and the bottom right panel shows the effects of Smad 7 overexpression on Smad 2/3 phosphorylation after TGF-β1 stimulation. (B). Immunofluorescence staining shows that Smad 2/3 nuclear translocation is affected by Smad 7. The top images are IF photos of Smad 2 (red) or Smad 3 (red); the bottom graphics show the statistical results for the nuclear density of Smad 2 and Smad 3. (C). The Western blot shows that the Smad 2/3 nuclear content is affected by Smad 7. The left part shows the data from the immunoblots; the right graphics show the quantitative analysis. (D). The effects of Smad 2, 3 and 4 on the phosphorylation of the other Smad protein. Smad 2, 3 and 4 was knocked down in fibroblasts mediated by the lentivirus. The right panel shows the knockdown efficiency of Smad 2, 3, 4 and 7.
