SKIP Downregulation Increases TGF-β1-Induced Matrix Metalloproteinase-9 Production in Transformed Keratinocytes
Figure 1
TGF-β1 induces MMP-9 expression and activates Smad3 and ERK1,2 signaling in transformed keratinocytes, PDV. (a) Zymography analysis of MMP-9 in serum-free conditioned media. PDV cells were treated for 24 h with different concentrations of TGF-β1; the media collected were subjected to electrophoresis on a gelatin-polyacrylamide gel. (b) RT-PCR analysis of MMP-9 expression. PDV cells were treated for 24 h with 5 ng/ml of TGF-β1, and mRNA obtained was reverse transcribed to cDNA. PCR products were separated in agarose gels. GADPH was used as housekeeper gene to verify the equal amount of cDNA in samples. (c) Western blot analysis for TGF-β1-induced activation of Smad3 and ERK1,2 MAPK. Cells were treated with 5 ng/ml of TGF-β1 at times indicated. The samples were assayed for phosphorylated forms of Smad3 and ERK1,2. The detection of total forms of Smad3 and ERK1,2 were used to confirm the amount of each protein loaded.
