For kinetic degradation assays, GSK3β-HiBiT KI HEK293 cells were reverse transfected into white bottom opaque 96-well assay plates. Briefly, GSK3β-HiBiT KI HEK293 cell suspension at a density of 2×10 6 per ml was prepared in standard culturing media. 10 mL of cell suspension mixed with transfection mix (1 mL of OptiMEM (Gibco) containing 3:1 ratio of FuGENE HD (Promega) and LgBit (Promega)). Then cells were seeded into 96-well assay plate as a 100 µL cell suspension per well. After 24 hours of transfection, cell media was removed and 90 µL of CO2-independent medium supplemented with 10% FBS (Gibco) and 1:100 dilution of Endurazine (Promega) and incubated for 2.5 hours at 37 ºC. Test compounds were added onto cells and luminescence signals were monitored and recorded on GloMAX discover plate reader (Promega) in kinetic mode every 5 minutes. Data were normalized to DMSO conditions and plotted over time and degradation rates were calculated via Michaelis-Menten non-linear fitting analysis using Graphpad Prism (v10.1.2).