Figure 4.

Real-time RT-PCR and western blot analyses for EMT-related molecules. (A) Real-time RT-PCR for EMT-related genes. To evaluate whether hypoxia mediates the EMT in H3122 cells, changes in the mRNA expression levels of EMT-related genes were evaluated using real-time RT-PCR. The experiment was performed in triplicate. Hypoxia upregulated SLUG mRNA expression (48 h, ^*P=0.030 and 96 h, ^*P=0.0053). E-cadherin mRNA expression was clearly downregulated under hypoxia (48 h, ^*P=0.035 and 96 h, ^*P=0.042). The expression of mesenchymal marker VIM and FN1 mRNA was also upregulated (48 h, ^*P=0.025 and ^*P=0.079, respectively and 96 h, ^*P=0.031 and ^*P=0.044, respectively). Columns, mean of independent triplicate experiments; error bars, SD; ^*P<0.05. (B) Western blot analyses for EMT-related molecules. Consistent with the mRNA changes, hypoxia time-dependently upregulated the protein expression of slug, fibronectin, and vimentin and downregulated the expression of E-cadherin in the cell line, along with HIF1α upregulation. β-actin was used as an internal control.