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. 2014 Sep 23;42(18):11589–11600. doi: 10.1093/nar/gku860

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Focal adhesion pathway gene vinculin as a direct transcriptional target of metastasis suppressor NME2. (A) Representation of NME2-target genes enriched in KEGG focal adhesion pathway; names are mentioned against highlighted boxes. Genes in blue color showed changes in mRNA level only (indirect targets); in comparison, genes with asterisk (*) showed alteration in mRNA level and binding of NME2 to their promoter as well (direct targets). (B) ChIP-PCR validation of NME2 occupancy of vinculin promoter: scheme showing NME2-motif within vinculin promoter; ChIP assay shows NME2 occupancy at the vinculin promoter; mock represents immunoprecipitation using non-specific isotypic IgG. (C) Over expression of NME2 represses (left panel), and stable depletion of NME2 enhances vinculin promoter activity in a luciferase reporter assay (right panel); 3- and 50-fold depletion corresponds to 66% and 98% decreased expression compared to control; all significance values: *P < 0.05, **P < 0.01; Student's t test, error bars represent standard deviation (SD).