Fig. 2: Screening of TEAD IPD library and hit identification.
From: Expanding the toolbox to develop IAP-based degraders of TEAD transcription factors
a IPD libraries were screened in a 20 h dose–response using a TEAD1 luciferase degradation assay (HiBiT-TEAD1, NCI-H2052 cells). Compounds with Dmax > 40% and DC50 < 1000 nM were further selected for a follow-up 2 concentration screen for endogenous TEAD1 degradation (20 h treatment with 0.3 and 3 µM IPD, NCI-H2052 cells). Three IPDs that achieved >40% degradation of endogenous TEAD1 were selected. b Chemical structures of the three IPDs selected (ALP A232, ALP2 A531, and XB2 A538) representing different IAP binder series, linkers and exit vectors (left panel), and corresponding endogenous TEAD1 and HiBiT-TEAD1 degradation results (middle and right panel). For endogenous TEAD1, NCI-H2052 cells were treated with IPDs (0.3 or 3 µM) or DMSO control for 20 h and RIPA lysates generated. Following capillary western electrophoresis, these were probed with TEAD1 and GAPDH (loading control) antibodies. Shown is one representative capillary western image for each IPD out of n = 2 independent experiments performed. Uncropped blot images are available in Supplementary Data 1. Percentage endogenous TEAD1 degradation (represented as mean % values) was calculated relative to 100% value of DMSO controls. For HiBiT-TEAD1 degradation (screening assay), NCI-H2052 cells stably expressing HiBiT-TEAD1 were treated for 20 h with a dose–response of IPDs or DMSO control. Percentage TEAD1 remaining was plotted based on HiBiT luminescence normalized to CTG (HiBiT/CTG ratio) relative to vehicle control. Plotted data represent individual data points from n = 3 independent experiments. Degradation DC50 and Dmax were fitted as described92.
