Fig. 2

Hypoxia-induced modulation of PTEN expression and EMT in H358 cells. a Cell extracts were analyzed for the levels of total and phosphorylated PTEN at the indicated time. A blot is representative of three independent experiments. b The ratio of p-PTEN to total PTEN is presented as an intensity level relative to that in H358 naïve cells at 0 min. Data were shown as the mean ± standard error (SE). The experiment was performed three times with similar results. Asterisk indicates p < 0.05. c Western blotting analysis for fibronectin and E-cadherin at the indicated time points was carried out. A blot is representative of three independent experiments. d The ratio of fibronectin to E-cadherin (F/E ratio) was evaluated, compared with that in cells treated under hypoxia at 0 min (bottom). Asterisk indicates p < 0.05. e, f By using confocal laser scanning microscopy and imaging software, we evaluated the fluorescence intensities of β-catenin (red) and E-cadherin (green) in the cells under normoxia or hypoxia. Hoechst33342 (blue) was utilized for nuclear staining. The left and right images in e shows cells cultured under normoxia and hypoxia, respectively. The upper and lower panels in f plot the fluorescence intensity of β-catenin red, E-cadherin green, and nucleus blue over a cross-section of cells cultured under normoxia and hypoxia, respectively. Data are representative of at least three independent experiments