Fig. 1: CeTEAM is predicated on stability-dependent biosensors to measure drug binding.

a U-2 OS V5-MTH1 G48E cells were treated with the indicated MTH1 inhibitors for 24 hours. b HCT116 3-6 3xHA-NUDT15 R139C cells were incubated with the indicated molecules for 72 hours. c U-2 OS PARP1 L713F-GFP cells were treated with PARP inhibitors for 24 hours. Biosensors were pre-induced with doxycycline, and all blots are representative from two independent experiments. d A schematic description of CeTEAM. Stability-dependent drug biosensors (blue) containing a destabilizing mutation (yellow) accumulate in the presence of binding ligand (pink) and detection can be facilitated by protein fusion tags (orange) to measure drug-target engagement. The presence of endogenous target protein (gray) and physiological conditions enable phenotypic multiplexing and discerning of on- from off-target effects of test compounds.