Fig. 2

CCL5 secreted by endothelial cells mediates AR decrease in CWR22Rv1 and C4–2 cells and enhances cell invasion. a Cytokine array analysis. CWR22Rv1 or C4–2 cells were cocultured with or without HUVEC cells for 2 days and the conditioned media collected for cytokine array analysis. b & c Cytokines that differentially secreted in conditioned media between two conditions were analyzed. d CCL5 ELISA assay for CCL5 level in conditioned media e. Quantitative PCR analysis of CCL5 in HUVEC. f CWR22Rv1 or C4–2 cells (1 × 10⁵/well) were treated with 20 ng/mL of human recombinant CCL5 (Peprotech) for 6 h, then western blot analysis was performed for AR expression. g CWR22Rv1 or C4–2 cells were transfected with MMTV-luc containing ARE. After transfection, cells were treated with 20 ng/mL of CCL5 for 24 h in the presence of DHT as indicated, and luciferase activity was measured. h & i C4–2 and CWR22Rv1 cells were cocultured with HUVECs in the presence of anti-CCL5–neutralizing antibody (R&D Systems) for 36 h, invasion assay or western blot analysis was conducted