Figure 2 | Cell Death & Disease

Figure 2

From: The embryonic transcription factor Brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies

Figure 2

Inhibition of Brachyury expression is associated with a mesenchymal-to-epithelial transition (MET) and decreased resistance to chemotherapy and radiation. H460 cells were stably transfected with vectors encoding for a nonspecific control shRNA (Control), or either of two different Brachyury-specific shRNAs (shRNA1 and shRNA2). Protein lysates collected from these cells were analyzed for expression of Brachyury, plakoglobin, fibronectin and vimentin by western blot (a) or by indirect immunofluorescence staining (b) for Brachyury, fibronectin, and vimentin. (Magnification × 40). (c) cDNA generated from H460 control.shRNA or Brachyury.shRNA-1 and -2 were analyzed for expression of Brachyury, Snail and Slug by quantitative real-time PCR, as compared with GAPDH. (d) Invasion assays were also performed with these cells. Indicated tumor cells were treated with various doses of chemotherapy (ng/ml) (e) or gamma radiation (Gy) (f) and assayed after 5 days culture for survival in comparison with untreated cells. Assays were performed in triplicate or quadruplicate; error bars correspond to S.E.M. Data shown are representative of at least two separate experiments (*P<0.05, **P<0.01, ***P<0.001)

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