Fig. 5. 1ae decreases interactions between AR and the transcription machinery.

a, Schematic of the method for small-molecule treatment of cells and the BioID experiment. b, BioID–MS of LNCaP MTID-AR-WT cells treated with EPI-001 (10 μM, 1 h), followed by treatment with DHT and biotin (2 h). Intensity data were obtained from SAINTq analysis. Shown is the log₂(FC) of the intensity of the interaction in inhibitor-treated versus DMSO-treated cells. Decreased interactors in inhibitor-treated cells relative to DMSO-treated cells are shown in blue; P < 0.05. The dashed lines mark where log₂(FC) < −1.5. Increased interactors in inhibitor-treated cells relative to DMSO-treated cells are shown in red; P < 0.05. The dashed lines mark where log₂(FC) > 1.5. Proteins of interest are annotated (n = 3). c, As in b, BioID–MS of LNCaP MTID-AR-WT cells. Cells were treated for 1 h with 1ae (5 μM) and then for 2 h with DHT and biotin (n = 3). d, (Total mean) TMean intensities of peptides identified by MS (n = 3) from SAINTq data of individual proteins (bait), total interactors (all) or collated known AR interactors sourced from BioGrid (https://thebiogrid.org/) were compared between LNCaP MTID-AR-WT cells treated with DMSO and those treated with small-molecule inhibitors. e, GO search terms of key biological processes and molecular functions in SAINTq intensity data from b and c (BFDR < 0.02, depleted = log₂(FC) < −1.5), obtained from the LNCaP MTID-AR-WT cells treated with small-molecule inhibitors versus DMSO. Full categories are available in Supplementary Data Table 2. f, BioID–MS of the LNCaP MTID-AR-WT interaction with Mediator complex in cells treated with small-molecule inhibitors versus DMSO. SAINTq data, color indicates strength of interaction change from logFC₁₀ TMean of intensity. g, The TMean intensity of interactions with the MED1 subunit of Mediator was compared between LNCaP MTID-AR-WT cells treated with DMSO or small-molecule inhibitors. SAINTq data were used. Statistical significance was determined by Student’s t-test against the control group. h, Results from a PLA in LNCaP MTID-AR-WT cells, using the indicated antibodies shown in cyan with DAPI staining shown in magenta. Cells were treated with small-molecule inhibitors or DMSO and DHT at the indicated times. Scale bars, 10 μm. i, Distributions of PLA foci per cell, each dot corresponds to a cell and boxes shown the mean and quartiles of all cells (n > 20 cells).

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