Fig. 2
From: Epithelial-to-mesenchymal transition in FHC-silenced cells: the role of CXCR4/CXCL12 axis
FHC-silencing and EMT gene expression in MCF7 cells. a E-Cadherin and Vimentin expression were evaluated by Real-time PCR, Western Blot and Immunofluorescence analysis. Real-time PCR were performed on total RNA extracted from MCF-7shRNA and MCF-7shFHC cells. Final results represent mean ± SD of three independent experiments. Statistical significance was evaluated by Student t-test (***, p < 0.001, ****, p < 0.0001). Western Blot analysis were performed on 50 μg of total protein extract from MCF-7shRNA and MCF-7shFHC cells. γ-Tubulin was used as loading control. For the Immunofluorescence analysis MCF-7shRNA and MCF-7shFHC cells were fixed and incubated with polyclonal anti-E-Cadherin or anti-Vimentin antibody (1:100) followed by incubation with the appropriate secondary antibody. Nuclei were visualized by DAPI staining. Images were collected using a Leica TCS SP2 confocal microscopy system (63X). Images are representative data from one of three experiments. b Western Blot analysis of ZEB1 was performed on 50 μg of total protein extract from MCF-7shRNA and MCF-7shFHC cells. γ-Tubulin was used as loading control. c Real-time PCR for Twist and Slug were performed on total RNA extracted from MCF-7shRNA and MCF-7shFHC cells. Final results represent mean ± SD of three independent experiments. Statistical significance was evaluated by Student t-test (**, p < 0.001)