Carboplatin

Cells expressing different HA-PKM2 mutants were seeded into 12-well plates, and the cell confluence area was monitored every 4 h over a 3-day period using an IncuCyte Live-Cell Analysis System (Essen Bioscience). To determine cell viability, cells transfected with or without CIP2A siRNA were plated into 96-well plates, and cytotoxic activity was assessed after the indicated drug treatment for 48 h using a Cell Counting Kit-8 (CCK-8; Vazyme) assay. The effects of the indicated compounds (Supplementary Table S3) on the cells were detected by CCK-8 assay, and the potential synergistic effect of the two compounds was calculated by the Bliss model using Combenefit software^{72 (link)}. For colony forming activity, the cells were plated into 12-well plates and treated with TEPP-46 and/or 2-DG for 48 h, and fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. To evaluate the effects of CIP2A overexpression on cell apoptosis, H1299 cells were transiently transfected with empty vector or Flag-CIP2A, and 24 h later the cells were treated with 0.3 mM carboplatin (APExBIO, Houston, TX, USA) for another 24 h. The cells were stained with Annexin V/FITC-propidium iodide Apoptosis Detection kit (APExBIO, Houston, TX, USA) and analyzed by flow cytometry (BD Biosciences, San Jose, CA, USA).