Figure S14. The levels of ubiquitinated and acetylated Slug. (a) To identify the ubiquitination sites of Slug, HEK293T cells were cotransfected with plasmids encoding different 3xFlag-tagged Slug mutants and Myc-tagged ubiquitin and were then treated with the proteasome inhibitor MG132 (10âÎźM for 6âh). Immunoprecipitation of the Slug protein using anti-Flag antibodies was analyzed by immunoblotting with anti-Flag antibodies. The asterisk and arrowhead indicate Slug modified and not modified by ubiquitin, respectively. (b) The acetylation of Slug. Different 3xFlag-tagged Slug mutants were transfected into HEK293T cells in the presence or absence of the sirtuin deacetylase inhibitor NAM and the HDAC inhibitor TSA. Acetylation of Slug was evaluated via immunoprecipitation and western blot analysis. Asterisk indicated acetylated Slug. (PDF 113 kb)