Fig. 4: Evaluating the degradation efficiency and ternary complex formation ability of BCL-xL/2 induced by WH244 and 753b, respectively.
From: Development and crystal structures of a potent second-generation dual degrader of BCL-2 and BCL-xL
a, b HiBiT degradation assay to measure the HiBiT-BCL-xL (a) and HiBiT-BCL-2 (b) degradation after 6 h treatment with different doses of WH244 or 753b. Data are represented as mean of two biological replicates. c, d HiBiT degradation assay to measure the time-dependent HiBiT-BCL-xL (c) and HiBiT-BCL-2 (d) degradation with two different doses of WH244 or 753b. Data are represented as mean of two biological replicates. e, f The formation of VCB/PROTAC/BCL-xL (e) and VCB/PROTAC/BCL-2 (f) ternary complexes in a cell-free condition was determined by AlphaLISA assay. Data represent the mean of a single experiment with 2 technical replicates. g, h Cellular ternary complex formation induced by 753b and WH244. 293T cells were transiently transfected with HiBiT-BCL-xL, LgBiT and HaloTag-VHL (g) or HiBiT-BCL-2, LgBiT and HaloTag-VHL (h) and then treated with a serial dilution of 753b or WH244 for 4 h. Cells were pretreated with proteasome inhibitor MG132 for 2 h to block the degradation of BCL-xL/2. Data are expressed as mean of three biological replicates. Source data are provided as Source data file.
