Additional file 1 of SIRT1 activation promotes energy homeostasis and reprograms liver cancer metabolism
Creators: name: Varghese, Benluvankar
name: Chianese, Ugo
name: Capasso, Lucia
name: Sian, Veronica
name: Bontempo, Paola
name: Conte, Mariarosaria
name: Benedetti, Rosaria
name: Altucci, Lucia
name: Carafa, Vincenzo
name: Nebbioso, Angela
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Chemical Sciences not elsewhere classified
subject: Cancer
subject: Computational Biology

description: Additional file 1: Figure S1. Cell proliferation upon treatment with SCIC2.1 at the indicated concentrations and times using xCELLigence. Figure S2. (a), In vitro enzymatic assay of SIRT1 performed on different SIRT1 modulators at the indicated concentrations, and counter-screening of NMase activity performed as in the SIRT1 assay. (b), In vitro SIRT3 enzymatic assay and counterscreening performed using different SIRT1 modulators at the indicated concentrations. (c), In vitro SIRT6 enzymatic assay performed using different SIRT1 modulators at the indicated concentrations. Statistical significance was calculated using the student t-test or one-way ANOVA using GraphPad Prism 9.4.0, and statistical significance is expressed as **p-value &lt; 0.01, ****p-value &lt; 0.0001. Error bars represent the standard deviation (SD) of three biological replicates. Figure S3. (a), cell cycle regulation of HepG2 cells glucose deprived for 24 h, 36 h, and 48 h, and graph showing cells in pre-G1 phase. (b), cell cycle regulation of HepG2 cells serum starved for 24 h, 3 6 h, and 48 h, and graph showing cells in pre-G1 phase. Statistical significance was calculated using the student t-test or one-way ANOVA using GraphPad Prism 9.4.0, and statistical significance is expressed as *p-value &lt; 0.05, ****p-value &lt; 0.0001. Error bars represent the standard deviation (SD) of three biological replicates. Figure S4. Mitochondrial respiratory parameters following glucose deprivation and bioenergetics parameters associated with SCIC2.1 treatment, and oxygen consumption rate (OCR) of HepG2 cells. (a), Glucose (25 mM) exposure followed by treatment with SCIC2.1. (b), Glucose (5 mM) exposure followed by treatment with SCIC2.1. Histograms show the different parameters (Basal Respiration, Proton Leak, Coupling Efficiency and Nonmitochondrial Oxygen Consumption). (c), extracellular acidification rate (ECAR) of HepG2 cells. Glucose (25 mM) exposure followed by treatment with SCIC2.1. (d), Glucose (5 mM) exposure followed by treatment with SCIC2.1. Histograms show the different parameters (Glycolytic Capacity and Glycolytic Reserve). Statistical significance was calculated using the student t-test or one-way ANOVA using GraphPad Prism 9.4.0, and statistical significance is expressed as *p-value &lt; 0.05, **p-value &lt; 0.01, ***P &lt; 0.001, nsp-value &gt; 0.05 vs control. Error bars represent the standard deviation (SD) of three biological replicates. Figure S5. Oil Red O staining of lipid droplet accumulation in HepG2 cells treated with SCIC2.1 (25 µM) and Ex-527 (5 µM) at the indicated time points. Images were acquired using a BioTeK Cytation 5 reader. Table S1. List of primers used in this study.