PKM2 Activity Assay

Pyruvate kinase activity was measured as previously described (27). Briefly, for cell line experiments, the medium was replaced with fresh medium 1 h prior to the start of treatment with DMAMCL. Where indicated, 100 μM pervanadate was added 10 min prior to cell lysis. Cells were lysed on ice with NP-40 buffer containing 2 mM DTT and protease inhibitors, then clarified by centrifugation at 12,000 rpm. Pyruvate kinase activity in lysates was determined by the pyruvate kinase activity assay kit (17). Results are represented as mean and s.e.m. of at least three independent experiments.