Fig. 3. AR interacts with EWSR1::WT1.

A Venn diagram of binding site overlaps between AR and EWSR1::WT1 in JN-DSRCT-1 cells identifies a set of 338 co-occupied sites. HOMER motif analysis on these co-occupied sites identified the WT1 motif as the most highly enriched motif and failed to identify the AR binding site as enriched. Venn diagram comparing DSRCT co-occupied sites with AR binding sites in the prostate cell line LNCaP identifies only 10 common binding sites. B DNA tracks of ChIP-seq data of EWSR1::WT1, AR, or input controls in JN-DSRCT-1 demonstrating co-occupied sites in FGFR4, CCL25, and MERTK. C Immunoprecipitation-Western blot using anti-AR or control antibody demonstrating an interaction between AR and EWSR1::WT1 as well as native EWSR1 in JN-DSRCT-1 and BER-DSRCT cells. D Immunoprecipitation-Western blot using anti-AR or control antibody in BER-DSRCT cells overexpressing FLAG tagged WT1 or EWSR1. E Venn diagram comparing the regulation of genes by EWSR1::WT1 (from RNA-seq data) and the closest gene to each genomic region co-occupied by EWSR1::WT1 and AR. F Genomic feature annotations of co-occupied sites that are associated with EWSR1::WT1 upregulated genes (UP), EWSR1::WT1 downregulated genes (DOWN) or genes not significantly altered by EWSR1::WT1 (STABLE). G RT-qPCR of four EWSR1::WT1 upregulated and co-occupied genes (MERTK, FGFR4, EPHB3, CCL25) in DSRCT cells treated with vehicle ctrl (CTRL), 1 nM DHT (DHT), 10 µM enzalutamide (ENZ), or 1 nM DHT and 10 µM enzalutamide (DHT + ENZ) (n = 2 independent samples).