Fig. 6.

SIRT1 nuclear activity maintains RAB25 repression stability. a–c Expression and localization of ZEB2 and SIRT1 in MCF7 upon ZEB2 induction (+dox) measured by a qRT-PCR and immunoblotting from b total protein extracts or c cytoplasmic (cyto) and nuclear (nucl) extracts. SIRT1 band density was measured and represented as histograms. Expression in control was set to 1. d, e RAB25 and CDH1 mRNA expression were measured by qRT-PCR d 24 h after ZEB2 induction (+dox) with SIRT1 inhibitor (EX-527, 1 µM) (+dox/EX-527) or e 24 h after ZEB2 induction and 48 h of doxycycline withdrawal (+dox) with SIRT1 inhibitor (EX-527, 1 µM) (+dox/EX-527). f Kinetic of RAB25 expression measured by qRT-PCR 24 h after ZEB2 induction and 24 or 48 h after doxycycline withdrawal (control) with SIRT1 inhibition (EX-527, 1 µM) (+EX-527). RAB25 level at control time point was set to 1, and s.d is shown. For all analyses, p values were determined using two-way ANOVA (**p < 0.01;***p < 0.001; ns nonsignificant). Three independent experiments were performed