Figure 5.

miR-151a reduces E-cadherin expression in NSCLC cells. (a) The relative expression levels of E-cadherin (left), fibronectin (middle) and Slug (right) were determined in stably miR-transduced A549 cells by RT-qPCR. (n=3 independent cell cultures and experiments, 3 technical replicates of each). (b) Representative confocal images of stably transduced A549s immunofluorescently stained with E-cadherin. Scale bar=10 μm. Relative fluorescence levels are quantified of the shown biological replicate (n=3 independent cell cultures and experiments, 12 technical replicates of each). (c) Immunoblot analysis of E-cadherin and tubulin protein levels in A549s transduced with miR constructs. Relative levels are quantified. One representative example of three is shown. (d) The expression levels of miR-151a was analyzed by in situ hybridization and compared with the expression of E-cadherin determined by immunohistochemical staining in three normal lung samples and 3 primary LACs (miR-151a expression: high=purple, low=light pink, E-cadherin expression: high=pink, low=blue). Cytokeratin 7 was included to identify cells of epithelial origin, for example, adenocarcinoma cells. Scale bar=50 μm. Throughout figure, all graphs are shown as mean±s.e.m. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 by two-tailed Student’s t-test. Uncropped version of blot is shown in Supplementary Figure S8.