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The absorbance of the treated and untreated cells was measured with a microplate reader (Bio-Rad) at 420�C480?nm. For the glucose-dependent cell death assay, cells were cultured with no-glucose DMEM?+ 10% FBS?+ galactose (4.5 g/l) or DMEM?+ 10% FBS?+ 2-Deoxy-D-glucose (2-DG, 10?mM) for 48?hr, and Dabigatran live/dead cells were quantified by cell counting with trypan blue exclusion and TC10 Automated Cell Counter (Bio-Rad). Data represent the mean?�� SD of three independent experiments. TS516 tumor sphere lines originated from a GBM patient at Memorial Sloan-Kettering Cancer Center, or U87 and U87-EGFRvIII cell lines were implanted into immunodeficient SCID/Beige mice for subcutaneous xenograft studies. SCID/Beige mice were bred and kept under defined-flora pathogen-free conditions at the Association for Assessment of Laboratory Animal Care-approved Animal Facility of the Division of Experimental Radiation Oncology, UCLA. For subcutaneous implantation, exponentially growing tumor cells in culture were trypsinized, enumerated by trypan blue exclusion, and resuspended at 3?�� 106 cells/ml in a solution of Dulbecco��s phosphate-buffered saline and Matrigel (BD Biosciences). Tumor growth was monitored with calipers by measuring the perpendicular diameter of each subcutaneous tumor. Tumors were treated with a PI3K/mTOR dual inhibitor (XL765; learn more 60?mg/kg) or normal saline every day for 19?days. Mice were euthanized if tumors reached 14?mm in maximum diameter or animals showed signs of illness. All experiments were conducted after approval by the Chancellor��s Animal Research Committee of UCLA. Unpaired Student��s t tests were performed unless otherwise noted. Error bars represented SEM unless otherwise noted, and statistical significance was indicated as ?p?Onalespib solubility dmso was kindly provided by Exelixis/Sanofi to I.K.M. This work is supported by grants from the National Institutes of Health (NS73831 and CA151819) (P.S.M.), NIH grant P01-CA95616 and a Fellow award from the National Foundation for Cancer Research (W.K.C.), and The Ben & Catherine Ivy Foundation and generous donations from the Ziering Family Foundation in memory of Sigi Ziering (T.F.C. and P.S.M.). ""Nonalcoholic fatty liver disease (NAFLD) is a key factor in the pathogenesis of type 2 diabetes (T2D) and affects one in three Americans (Petersen et?al., 2005?and?Samuel and Shulman, 2012).