Fig 2: Deacetylation destroys pro-glycolytic function of β-catenin. (A) IGV snapshot of transcriptome analysis of MYC between HCT116-WT and HCT116-CTNNB1 KD cells. (B) IGV snapshot of transcriptome analysis of MYC between wild-type and SIRT1 overexpression tumor cells. (C) % of β-catenin interaction sites in genomes of HCT116 and DLD1 cells with SIRT1 overexpression. (D) Immunoblot analysis of glycolysis-related genes in malignant cells treated with control Flag vs. Flag-SIRT1. (E) Immunoblot analysis of glycolysis-related genes in malignant cells treated with control shRNA vs. SIRT1 shRNA. (F) ECAR was examined in HCT116 and DLD1 cells treated with 20 μM EX527. Each assay consisted of 3 wells each for the treatment and control groups. Data are means ± SEM of three independent assays. **P < 0.01, ***P < 0.001. (G) CTNNB1 KD HCT116 cells and DLD1 cells transfected with GFP-tagged CTNNB1 WT, mutant K49Q and mutant K49R. Then the subcellular localization of transfected β-catenin was examined by confocal microscope. Scale bar, 10μm. (H) The fluorescence intensity of GFP in nuclei of cells in (G) were determined using ImageJ. The number of cells used in statistics for each cell type ranged from 8-10. ***P < 0.001. (I) Analysis of key glycolytic genes expression in CTNNB1 KD HCT116 cells and DLD1 cells transfected with CTNNB1 WT, mutant K49Q and mutant K49R respectively. (J) HCT116 CTNNB1 KD were transfected with CTNNB1 WT, mutant K49Q and mutant K49R. ECAR were examined by Seahorse XFp extracellular flux analyzer. Each assay consisted of 3 wells each for the treatment and control groups. Data are means ± SEM of three independent assays. WT-CTNNB1 was regarded as reference. *P < 0.05, **P < 0.01, ***P < 0.001. (K) Colony-forming efficiencies were determined for HCT116 and DLD1 cells treated with 10 μM XAV939. ***P < 0.001. (L) The tumor suppression effect of 10 mg/kg XAV939 was evaluated in nude mice subcutaneously injected with DLD1 cells (1 × 106/mouse, n = 8/group). From day 12 after the initial injection, tumor volumes were measured with a vernier caliper every 3 d. Tumor tissues were collected and weighed at the end of the assay. ***P < 0.001, ns: nonsignificant.