Metabolic Profiling of Drug-Treated Cells

For lactate, LDH activity, NAD+/NADH and NADP+/NADPH measurements, cells were harvested at various time points following drug treatment using appropriate buffers and frozen in liquid nitrogen. Intra-cellular lactate, NAD+, NADH, NADP+ and NADPH levels along with ex vivo LDH activity were analyzed using commercially available colorimetric assays (BioVision, CA, USA), according to the manufacturer’s instructions as previously described by our group and others^{17 ,19} . Intra-cellular ATP levels were measured using commercially available luminescent assays (Promega, WI, USA) as previously described by our group and others; ATP levels were normalized to cell number quantified using Hoechst staining as previously described by our group^{18 (link)}.