Additional file 6: of Hypoxia-induced Slug SUMOylation enhances lung cancer metastasis

Figure S6. Characterization of Slug and Slug5M protein. (a) The DNA-binding ability of Slug is not altered by the inserted mutations. Equal amounts of in vitro-translated Slug and Slug5M were used in the EMSAs (left panel). Slug and Slug5M bound to the E-box C probes in a dose-dependent manner (+: 0.1 μl; ++: 0.3 μl; +++: 1 μl) (right panel). Anti-Slug antibodies were used to confirm that the shifted bands were formed specifically by Slug and Slug5M. (b) The protein stability of Slug is not altered by the inserted mutations. Protein stability was not significantly different between the wild-type and mutant forms of Slug. Slug- and Slug5M-overexpressing HEK293 cells were treated with cycloheximide (CHX) to prevent further protein synthesis for the indicated periods. The expression of Slug was analyzed by immunoblotting. β-actin was used as the internal control. Relative densitometry results are plotted in the bottom panel. (PDF 68 kb)