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PubMed Central, Figure 4: Nat Cell Biol. 2012 Nov; 14(11): 1212–1222. Published online 2012 Oct 21. doi:  10.1038/ncb2607
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Nat Cell Biol. Author manuscript; available in PMC 2013 May 1.
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Nat Cell Biol. 2012 Nov; 14(11): 1212–1222.
Published online 2012 Oct 21. doi:  10.1038/ncb2607

Figure 4

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Overexpression of Elf5 reverses mesenchymal characteristics of MDA-231 cells

(a) Western blot analyses of HA-Elf5 and other EMT markers in MDA-231 cells transduced with vector (control) or HA-Elf5. Uncropped images of blots are shown in Supplementary Fig. S9. (b) Phase contrast images of parental, control or HA-Elf5-overexpressing MDA-231cells. Size bar = 100 μm. (c) Immunofluorescence analysis of control and HA-Elf5-overexpressing MDA-231 cells stained for the indicated proteins. Arrows mark the relocalization of nuclear β-CATENIN to the membrane and cytoplasm, restoration of circumferential F-actin belts, membrane localization of ZO-1 and adherens junctions, and reduced expression of VIMENTIN in Elf5-overexpressing cells. Size bar = 20 μm for β-CATENIN, and F-ACTIN, 60 μm for E-CADHERIN, and 40 μm for ZO-1 and VIMENTIN. (d, e) Quantitative RT-PCR analysis of the expression of EMT-related genes in control or HA-Elf5-overexpressing MDA-231 cells at 12 days (d) or 48h post-infection (e). Real time values were normalized to the housekeeping gene GAPDH. Experiments were performed three times, each with qRT-PCR in technical duplicate, and data presented as the mean ± SD. * p < 0.05 by Student’s t-test. (f) Transwell migration assay of MDA-231 cells with or without Elf5 overexpression. The data are shown as mean ± SD collected from 10 fields of 3 independent experiments. Student’s t-tests were performed to assess statistical significance. (g, h) Matrigel invasion assay of MDA-231 cells with or without Elf5 overexpression. Two time points were observed for invasion assays (24 h in g and 48 h in h). The data were collected from 10 fields, performed in triplicate, and shown as mean ± SD. * p < 0.05 by Student’s t-test.

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