ON-TARGETplus siRNA pools for gene expression knockdown.

List of the 44 3D structures of HSC70 (HSPA8) in complex with the BAG domain of BAG-1 used in comparative structural analyses.

Antibodies used for western blot analyses.

Development and characterization of BAG-1 knockout mice.

Synthego BAG-1 sgRNA for BAG-1 CRISPR knockout clones.

Development of castration resistant prostate cancer patient derived xenograft organoids.

Thio-2 reduces genome-wide androgen receptor binding in LNCaP prostate cancer cells.

Mouse organ hematoxylin and eosin, and BAG-1 immunohistochemistry, in BAG-1 knockout mice.

Mouse organ hematoxylin and eosin, and BAG-1 immunohistochemistry, in BAG-1 knockout mice.

TaqMan probes for qRT-PCR analyses.

Thio-2 downregulates C-MYC expression through a BAG-1 independent mechanism.

BAG-1 knockout female mice are fertile and viable with reduced gestation.

Treatment with Thio-2 inhibits transactivation of the unstimulated and stimulated androgen receptor and the constitutively active androgen receptor splice variant-7.

Thio-2, may bind the N-terminus of the androgen receptor through a similar binding mode to EPI-001.

Description of Materials and Methods not contained within the main Materials and Methods section

Protocols and antibodies for immunohistochemical analyses.

In-vivo treatment of patient derived model of lethal prostate cancer demonstrates Thio-2 to be well tolerated with associated impact on AR signaling and growth.

Thio-2 downregulates androgen receptor signaling and inhibits growth of prostate cancer cell lines through a BAG-1 independent mechanism.

Androgen receptor (AR) splice variants, of which ARv7 is the most common, are increased in castration-resistant prostate cancer, but the extent to which they drive AR activity is unclear. We generated a subline of VCaP cells (VCaP16) that is resistant to the AR inhibitor enzalutamide (ENZ). AR activity in VCaP16 is driven by ARv7, independently of full-length AR (ARfl), and its cistrome and transcriptome mirror those of ARfl in VCaP cells. ARv7 expression increases rapidly in response to ENZ, but there is a delay in gaining chromatin binding and transcriptional activity, which is associated with increased chromatin accessibility. AR and nuclear factor I (NFI) motifs are most enriched at more accessible sites, and NFIB/X knockdown greatly diminishes ARv7 function. These findings indicate that ARv7 can drive the AR program but that its activity is dependent on adaptations that increase chromatin accessibility to enhance its intrinsically weak chromatin binding.