Colony Formation Assay for PC-3 and LNCaP Cells

PC-3 and LNCaP cells with different treatment were resuspended in fresh medium and then seeded into the 6-well-plates (1000 cells/well). The cells were grown under the condition of 37 °C and 5% CO₂ for 14 days, and the medium was replaced every three days. Cells were fixed by 4% paraformaldehyde for 20 min, and stained using Giemsa Stain Kit (Abcam, ab150670) for 20 min at room temperature. Colony morphology were photographed and colony number was counted under Leica AM6000 microscope [41 (link)]. The colony formation assay was conducted with three independent biological replicates.