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. Author manuscript; available in PMC: 2024 Apr 2.
Published in final edited form as: Mol Cancer Ther. 2023 Oct 2;22(10):1166–1181. doi: 10.1158/1535-7163.MCT-23-0196

Figure 5. MPC309 regulates AR interactions with coregulatory proteins on chromatin.

Figure 5.

(A) Mammalian two-hybrid assay to detect the interaction between the AR LBD and the LxxLL motif. The Gal4-DBD fused to SRC1a 1241–1441 and VP16AD-LBD WT were co-transfected with the 5x Gal4 RE-Luc reporter (n=3). An unpaired t-test was used to test for statistical significance between the two groups and the difference was considered significant for P values <0.05 (**P <0.01, ***P<0.001). (B) Evaluation of the MPC309 effect on AR coactivator peptide recruitment, shown as a TR-FRET emission ratio. This experiment was performed using the LanthaScreen® TR-FRET AR Coactivator Assay PolarScreen™ Kit in agonist mode (n=2). An unpaired t-test was used to test for statistical significance between the two groups. (C) LNCaP-abl cells were cultured under androgen deprivation and treated with vehicle (DMSO) or 10nM DHT or 1μM MPC309 overnight. RIME was performed to identify co-regulators associated with AR on chromatin. Venn diagrams show the number of unique and common proteins associated with AR in Vehicle, DHT, and MPC309-treated cells. (D) Left: Top 25 proteins enriched in MPC309 compared to DHT-treated cells. Right: Top 25 proteins enriched in DHT compared to MPC309-treated cells. Fold-change was calculated by adding +5 to PSM counts, subtracting IgG, and normalizing to AR count in each sample. Common background proteins were removed from the analysis.