ADQ activates the Akt/GSK3β-mediated proteasome degradation of β-catenin in breast CSCs. (A) The expressions of β-catenin, ABCG2, P62 and LC3 in MDA-MB-231 CSCs were examined by western blotting after the indicated treatment. (B) The fractional or total expressions of β-catenin, ABCG2, P62 and LC3 in MDA-MB-231 CSCs were assayed by western blotting after the indicated treatment. (C) Representative immunofluorescent images of β-catenin in MDA-MB-231 CSCs spheres and its re-attached CSC cells. All values represent the means ± SD (n = 3, *p < 0.05, **p < 0.01 vs. Control group; #p < 0.05, ##p < 0.01 vs. Taxol group). (D) The impacts of ADQ on the β-catenin proteasome degradation pathway were evaluated by western blotting. The upper graph shows that the action of ADQ on β-catenin degradation was blocked by the proteasome inhibitor MG132. The lower graph indicates the influence of ADQ on β-catenin degradation was accelerated by the protein synthesis inhibitor CHX. All values represent the means ± SD (n = 3, *p < 0.05, **p < 0.01 vs. Control group; #p < 0.05, ##p < 0.01 vs. Taxol group). (E) Western blotting analysis demonstrated that ADQ administration notably affected the expressions of the phosphorylation of β-catenin, total/phosphorylation of GSK-3β as well as total/phosphorylation of AKT in a time- and dose-dependent manner. (F) Western blotting analysis showing the synergsitic effects of ADQ with either GSK-3β inhibitor LiCl or AKT inhibitor LY294002.
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