Fig. 3.

Rk1 and Rg5 inhibit TGF-β1-induced EMT. A. Chemical structure of Rk1 and Rg5. A. Chemical structure of Rk1 and Rg5. B. The antiproliferative effects of Rk1 and Rg5 on cell proliferation were determined by CCK-8 assay. A549 cells were treated with Rk1 or Rg5 at concentrations of 0 (DMSO was used as a control), 20 μM, 30 μM, 40 μM, 50 μM, and 60 μM and 0 μM, 100 μM, 150 μM, 200 μM, 250 μM, and 300 μM for 24 h or 48 h. C. Control A549 cells treated with DMSO appears to be of original epithelial morphology. TGF-β1 treatment induces mesenchymal morphology. However, A549 cells treated with Rk1 at concentrations of 45 μM and 50 μM of Rk1 or Rg5 at concentrations of 200 μMand 250 μM plus 5 ng/mL of TGF-β1 for 48 h show the epithelial phenotype. D and E. Changes in protein of the epithelial marker, E-cadherin and the mesenchymal marker vimentin were analyzed by Western blot analysis and densitometric quantification. β-Actin was used as a loading control. F. Rk1 or Rg5 increased E-cadherin expression and decreased vimentin expression in TGF-β1-stimulated A549 cells. The expression of E-cadherin and vimentin were detected by using immunofluorescence analysis with primary anti-E-cadherin and anti-vimentin. A549 cells were cultured in the TGF-β1 with Rk1 or Rg5 or without Rk1 or Rg5 for 48 h. Images show nucleus stained by (DAPI). ∗∗p < 0.01, ∗p < 0.05 versus control and ##p < 0.01 #p < 0.05 versus TGF-β1 treated control. The data are expressed as mean ± SD for triplicates.