Cell Viability Assay with Enzalutamide

C4-2, SaOS-2, and SaOS-LM2 cells lines were harvested after reaching 80% confluence, and 10⁵ cells were plated in 6-well plates for 24 h. C4-2 was used as a positive control for the expression of AR. Cells were then treated with enzalutamide (Selleck Chemical LLC, Houston, TX, USA, Cat.# S1250) at fold dilutions from 100 to 3 µM. In addition to enzalutamide treatment, untreated and methanol-treated (positive control) conditions were also tested. Each condition was performed in triplicate. Cells were incubated in their respective conditions for 72 h, after which they were harvested and counted for viability with Trypan blue (Bio-Rad, Cat.# 1450022).