Darolutamide daro

The following chemicals were used at concentrations indicated in the results and figure legends: dimethylsulfoxide (DMSO; Cat# D2650, Sigma Aldrich, Vienna, Austria), doxycycline (Dox; Cat# D9891, Sigma Aldrich), mifepristone (RU486; Cat# S2606), dexamethasone (Dex; Cat# S1322), prednisolone (Pred; Cat# S2570), apalutamide (Apa; Cat# S2840) (Selleck Chemicals, Munich, Germany), enzalutamide (Enza; Cat# HY-70002), abiraterone (Abi; Cat# HY-75054) (Hycultech, Beutelsbach, Germany), darolutamide (Daro; Cat# HY-16985), bicalutamide (Bic; Cat# HY-14249) (MedChemExpress, Stockholm, Sweden), docetaxel (Doc; Cat# MCE-HY-B0011), and cabazitaxel (Cab; Cat# MCE-HY-15459) (THP Medical Products).

LNCaP, LNCaP-GFP, LAPC4, LAPC9-GFP and 22Rv1 cells (2000 cells/well) were seeded in a 384-well black plate (Greiner Bio-One North America Inc., Monroe, NC, USA) in RPMI-1640 with 10% charcoal-treated FBS (FBS-CT, Wisent) and dihydrotestosterone (DHT) (Toronto Research Chemicals, Toronto, ON, CA) at 1 nM. The cells were then transduced with either CMV-TSTA (1 × 10⁴ infectious viral particle (IVP)) or PSEBC-TSTA (1 × 10⁵ IVP) adenoviruses. Bioluminescent imaging was performed as described below by using LV200 bioluminescence microscope. Media were replaced with appropriate treatments (DHT (1 nM), DHT + 10 μM Enzalutamide (Enza, MedChem Express, Monmouth Junction, NJ, USA), DHT + 10 μM Bicalutamide (Bica, Sigma-Aldrich, Oakville, ON, Canada), DHT + 10 μM Apalutamide (Apa, MedChem Express) DHT + 10 μM Darolutamide (Daro, MedChem Express)). Forty-eight hours later, bioluminescence imaging was repeated on the same cells. D-luciferin (3.5 mM) was added 20 min before each imaging time-point.