Myc reporter kit

The c-Myc activity was assessed using the Myc Reporter kit (BPS Biosciences) and the Dual-Luciferase Reporter System (Promega) according to the manufacturer’s instructions. Briefly, 100 μL (1.5 × 10⁵ cells/mL) of wildtype and Pkm2^−/− KCs were seeded into 96-well plates. After overnight incubation, when cells reached ~50% confluency, 1 µL of Reporter A (60 ng/µL) was transfected into cells using Turbofectin 8.0. After 48 h, cells were lysed in 25 µL Passive Lysis Buffer (provided in the Dual-Luciferase Reporter kit). 20 µL of cell lysate was transferred to 96-well plates and placed in a 96-well microplate luminometer (GloMax-Multi, Promega). 100 µL Luciferase Assay Reagent II and 100 µL Stop & Glo Reagent (both provided in the Dual-Luciferase Reporter kit) were sequentially injected, and firefly and Renilla luciferase activities were automatically measured. c-Myc activities were determined by the ratios of firefly to Renilla luciferase activities.

The c-Myc activity was assessed using the Myc Reporter kit (BPS Biosciences #60519) and the Dual-Luciferase Reporter System (Promega #E1910) according to the manufacturers’ instructions. c-Myc activities were determined by the ratios of firefly to Renilla luciferase activities.