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Figure S1: Overall survival probability of the patients according to NAMPT levels segregated by colon cancer stage. Figure S2. Tumorigenic properties induces by NAMPT Figure S3. Effect of a second shRNA against NAMPT in the tumorigenic capability of tumor cells. Figure S4. CD133+ single cell derived tumorspheres of HCT116. Figure S5.Tumorigenic capabilities NAMPT-dependent in HCT 116 +/+ p53 wild type. Figure S6. (A) GSE34053 database, from human colon tumors, have transcription profiles of sorted CD133+ cells. We analyzed the correlation of CD133+ cells with NAMPT expression along with the following: HES1, OCT4 and SOX9. (B) GSE14774 database includes transcription profiles of a supplementary colon cancer cell line, HT-29, both in monolayer culture and tumorspheres. We analyzed the correlation of NAMPT on either monolayer or tumorspheres along with the following: HES1, OCT4 and SOX9. (C, left graph) Clonal heterogeneity analysis reveals that 1 mM NMN can rescue the number of holoclones up to vector levels in both cell lines. (C, right graph) Clonal heterogeneity analysis reveals that treatment with 60 ng/mL visfatin results in the same effect over holoclones. Figure S7. Effect of the addition of the NAMPT product NMN, or the extracellular NAMPT (visfatin) to HCT116 p53 WT cells depleted of NAMPT. Figure S8. (A) Clonal heterogeneity analysis reveals that 2.5 nM FK866 abruptly decreases the number of holoclones in both cell lines. (B) Clonal heterogeneity analysis reveals that 1 µM sirtinol also abruptly decreases the number of holoclones. Figure S8C-I. Effect of the addition of the Sirt1 inhibitor Sirtinol to HCT116 p53 WT cells overexpressing NAMPT. Left half of the figure: we added 1 µM sirtinol to cells overexpressing NAMPT (NAMPT+S, Â-�, orange bars) and compared different properties to parental expressing only vector (v, Â-�, grey bars), shRNA only (sh, O, white bars) or NAMPT overexpression only (NAMPT, Â- , black bars). (C) growth curve, (D) clonogenic assay, (E) CD133 percentage of cells, (F) number of tumorspheres, (G) Colony type, (H) stem cell pathway effectors and (I) EMT effectors [*, p |
