Figure 4. GR inhibition resensitizes TLE3^KO cells to enzalutamide treatment.

(A) Western blot showing protein expression levels of TLE3 and GR in control and TLE3^KO cells cultured vehicle or enzalutamide (B) Coverage profiles for TLE3 and AR binding at the GR locus. (C) Long-term growth assay (14 days) showing the drug resistance phenotype of control and TLE3^KO cells with and without GR knockdown in the presence of vehicle or enzalutamide. (D) Western blot analysis for TLE3 protein levels in control and TLE3^KO cells shown in C, using GAPDH as a loading control. (E) mRNA levels for GR in control and TLE3^KO cells carrying shSCR or shGR constructs, shown in C. (F) Long-term growth assay (14 days) for cells harboring a control sgRNA or TLE3-targeting sgRNA cultured in the presence of vehicle, enzalutamide, mifepristone or the combination at indicated concentrations. (G) ChIP-qPCR showing GR occupancy at enhancers proximal to indicated genes. All samples were cultured in the presence of 10 μM enzalutamide with or without 1 μg/ml hydrocortisone (HC) as indicated.

Figure 4—figure supplement 1. GR-mediated gene regulation in TLE3^KO cells.

(A) Coverage profiles for TLE3 or H3K4me1 at the GR locus in LNCaP and LREX’ cells. (B) Core consensus sequences for AR and GR. (C) Quantification of long-term colony formation assays showing the functional phenotype of control and TLE3^KO, with or without GR knockdown, in cells treated with vehicle or enzalutamide. (D) ChIP-qPCR for H3K27 acetylation in control and TLE3^KO cells at indicated loci. (E) Overview of the genes listed in Figure 2B with GR-regulated genes (as shown by Arora et al., 2013) highlighted in green.