Smad2/3 Antibody - #AF6367

Figure 4 FOXC2 driving EMT changes in PMCs may be achieved through the TGF-β1-Smad2/3 signaling feedback loop. A,B) Western blot analyses of FOXC2, Smad2/3, and p-Smad2/3 protein levels among different groups. n = 3 per group. Compared with the vector group, **p < 0.01, *p < 0.05. Compared with the vector group in the presence of TGF-β1, #p < 0.05. C,D) Western blot analyses of Smad2/3 and p-Smad2/3 protein levels among different groups. n = 3 per group. Compared with the vehicle group, **p < 0.01. Compared with the FOXC2-KD group, ##p < 0.01. E) Immunofluorescence staining images of nuclear accumulation of TGF-β1-induced Smad2/3 (red) in FOXC2-KD PMCs. Scale bar = 10 µm. F,G) Western blot analyses of Smad2/3 protein abundance in the cytoplasm and nucleus among different groups. n = 3 per group. Compared with the vehicle group, **p < 0.01. Compared with the TGF-β1, ##p < 0.01. H) The cell viability of PMCs treated with different concentrations of DGM (5, 10, 20, 40, 80, and 160 µM) for 24 h by CCK8 assay. n = 12 per group. Compared with the vehicle group, **p < 0.01. I) The cell viability of PMCs treated with TGF-β1 and different concentrations of DGM (10, 20, and 40 µM) for 24 h by CCK8 assay. n = 10 per group. Compared with the vehicle group, **p < 0.01. Compared with the TGF-β1 group, ##p < 0.01. J) Western blot analyses of the protein abundance of FOXC2, α-SMA, E-cadherin, Smad2/3, and p-Smad2/3 in PMCs treated with TGF-β1 and different concentrations of DGM (10, 20, and 40 µM) for 24 h. K–M) qRT-PCR analysis of mRNA expressions of FOXC2, α-SMA, and E-cadherin in PMCs treated with TGF-β1 and different concentrations of DGM (10, 20, and 40 µM) for 24 h. n = 6 per group. Compared with the vehicle group, **p < 0.01. Compared with the TGF-β1 group, ##p < 0.01, #p < 0.05.

Figure 6. SCs promote the migration of fibroblasts and keratinocytes. (A) In vitro wound-healing assays of 3T3 cells cultured in mediums from 3T3, RSC96 or S16 cells. (B) Migration rates were evaluated according to the closure area at 18 h after lesion induction. (C) In vitro wound-healing assays of HaCaT cells cultured in mediums from HaCaT, RSC96 or S16 cells. (D) Quantification of wound closure at 18 h after scratching. (E) In vitro wound-healing assays of 3T3 cells cultured in medium from RSC96 or S16 cells after control or Tgfb3 RNAi (siCtrl or siTgfb3) treatment. (F) Quantification of wound closure of 3T3 cells under different conditions. (G) In vitro wound-healing assays of HaCaT cells cultured under the indicated conditions. (H) Quantification of wound closure in HaCaT cells under the indicated conditions. (I) Immunoblot analysis of the expression of aSMA, phosphorylated Smad2/3 (p-Smad2/3) and Smad2/3 in 3T3 or HaCaT cells after incubation with mediums from RSC96 or S16 cells for 24 h. The images presented are representative of three independent experiments. Scale bars: a, c, e, g, 200 mm. Unpaired t test. The data are presented as the mean ± SEM, *P