Figure 5

The EMT-inducing culture conditions increase miR-21 expression in mouse lung alveolar epithelial cells. (A) Representative flow cytometric analyses for the expression of pro-SFTPC in isolated mouse alveolar type 2 cells. Note that most of the isolated cells were positive for both EpCAM and pro-SFTPC. (B) Representative immunofluorescence staining for pro-SFTPC in the isolated cells. Most of the isolated cells were positive for pro-SFTPC (left), whereas EPCAM-CD45- cells were negative for pro-SFTPC (right). Scale bars: 20 μm. (C) The EMT-inducing culture conditions (EMT; cultured on fibronectin with TGF-β for 6 days) decrease lung epithelial cell marker gene expression and increase EMT marker gene expression in mouse lung alveolar epithelial cells. The relative expression of mRNA of lung epithelial cell marker genes (E-cadherin and SFTPC) or EMT marker genes (vimentin, α-SMA, Zeb1 and Zeb2) is shown. The values are mean ± SEM (n = 6). mRNA is expressed relative to levels observed prior to culture. The data were analyzed by one-way analysis of variance with a post hoc test (Scheffé’s test). * p < 0.05 versus cells not cultured (NC). † p < 0.05 versus control cultures (Con; cultured on Matrigel/collagen for 6 days). (D) The relative expression of microRNAs is shown. The values are mean ± SEM (n = 6). microRNA expression was relative to the levels observed prior to culture. The data were analyzed by one-way analysis of variance with a post hoc test (Scheffé’s test). * p < 0.05 versus NC. † p < 0.05 versus Con.