Ml265

The AMPK activator A769662 was purchased from APExBIO. The AMPK activator metformin, the mTORC1 inhibitor rapamycin and the metabolite succinate were obtained from Sigma-Aldrich. The fatty acid synthase (FAS) inhibitor C75 and the AMPK inhibitor Compound C were purchased from Cayman Chemical. The PFKFB3 inhibitor 3PO was from EMD Millipore. The human NMT1 cDNA ORF Clone and the appropriate control were purchased from Origene. Human NMT1 siRNA and control siRNA, human AMPKα siRNA and control siRNA, the pyruvate kinase M2 (PKM2) activator ML265, the PKM2 inhibitor Shikonin, the metabolites pyruvate and malic acid, were bought from Santa Cruz Biotechnology. Nucleofection kits from Lonza were used for cell transfections. Lysosomes were isolated with the Lysosome enrichment kit for cultured cells (Thermo Fisher Scientific). Concentrations of cellular free fatty acids were determined with the Free Fatty Acid Quantification Assay Kit (Abcam). All reagents were used according to the manufacturers’ instructions.