Fig. 1 |. Development of a high-throughput assay for evaluating off-target ZF degradation of pomalidomide-based PROTACs.

a, An automated imaging screen for the degradation of ZF degrons by pomalidomide analogues and pomalidomide-based PROTACs. Briefly, U2OS cells stably expressing 14 ZF degrons fused to eGFP were treated with PROTACs followed by imaging to assess ZF degradation. Decrease in the eGFP signal is the measure of ZF protein degradation. b, Degradation of validated and pomalidomide-sensitive ZF degrons inside cells by reported PROTACs in a dosing range of 4.3 nM to 6 μM. As shown in the schematic, each rectangle in the heatmap represents a single dose. Chemical structures with rectangle boxes and oval shapes indicate linker chemotype and target protein binding ligands respectively. Data are the mean of at least three independent replicates.