The SNAG and SLUG domains of Snail2 are required for efficient repression of <i>E-cadherin</i> promoter.

<p>(A) Schematic representation of the organization of mouse Snail2 protein. Upper, the different domains are represented with a color code: the SNAG domain, 1–9 amino acids (orange) the SLUG domain (96 to 123 amino acids) (green) and the five zinc fingers (blue to magenta) constituting the DNA binding domain (DBD). Middle and bottom, schematics of the ΔSNAG and ΔSLUG mutants with indication of the specific amino acids deleted. (B) The repressor activity of Snail2-HA wild type and the indicated mutants on the mouse <i>E-cadherin</i> promoter was analyzed on HEK293T cells. Reporter assays were performed with 100 ng of the indicated vectors as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036132#s4" target="_blank">Material and Methods</a>, and relative luciferase units (RLU) normalized to the activity obtained in the presence of a void control pcDNA3 vector. Results show the mean of triplicate experiments, performed on quadruplicate samples, +/− s.d. *p<0.05; ***p<0.001. Upper insets show western blot controls for equal expression of the different Snail2 forms. (C). Nuclear localization of Snail2-HA and the SNAG and ΔSLUG mutants in transiently transfected HEK293T cells as determined by immunofluorescence analysis.