Fig. 6: Specificity of endogenous TEAD degradation by IPDs.
From: Expanding the toolbox to develop IAP-based degraders of TEAD transcription factors
a Endogenous TEAD1 and TEAD4 degradation profiling in NCI-H2052 cells with 20 h treatment with ALP2 IPD A531, XB2 IPD A538 and ALP2 IPD A536. With compound concentrations represented in increasing concentration from DMSO to 10 µM, Top panel shows representative capillary western blot of TEAD4 and loading control GAPDH; middle panel, blot of TEAD1 and GAPDH and bottom panel shows degradation dose response curves with each data point representing mean ± SD of n = 2 biologically independent experiments, except for n = 5 for A531 (TEAD1) and n = 3 for A538 (TEAD1). Uncropped blot images are available in Supplementary Data 1. b HiBiT assay measuring Dmax of ALP2 IPD A531, XB2 IPD A538 and ALP2 IPD A536 after 18 h treatment of HiBiT-TEAD1–4 NCI-H226 transgenic cell lines, normalized to CTG viability assay. Compound dTAG-13 (heterobifunctional degrader of the FKBP12F36V sequence incorporated in the TEAD1–4 transgenic constructs)97 was used as positive control. Data are representing mean ± SD of n = 4 biologically independent experiments, each with two technical replicates. c Global proteomic analysis of XB2 hit A538 specificity in NCI-H2052 cells, treated with compound A538 (0.5 µM, 16 h) or DMSO (n = 5 biological replicates). Volcano plots show relative protein abundance (log2 fold change) vs significance (−log10 p-value) of quantified proteins. Proteins significantly altered lie above the horizontal dashed line (adjusted p-value “or” FDR ≤ 0.05) and beyond vertical cut off lines (left, 1.25 times downregulated; right, 1.25 times upregulated in A538 treated cells). A complete protein list is provided in Supplementary Data 2.
