Inhibition of AhR signaling decreases growth of prostate cancer cells.

<p><b>A.</b> Cells were grown in a 96 well plate at 5.0×10³ cells per well. The cells were treated with DMSO or 1 µM, 5 µM, 10 µM, 25 µM or 50 µM of CH223191. Cell growth was measured using Promega CellTiter 96 Cell Proliferation Assay per manufactures instructions. <b>B.</b> Confirmation of AhR protein expression in DU145 cells with scrambled vector control (SCR) and shRNA lentivirus against AhR (-AhR) by western blotting. 20 µg of protein isolated from was analyzed by western blotting. β-action is used as a loading control. Proliferation of DU145(SCR) and DU145(-AhR) cells were analyzed using the CellTiter 96 Cell Proliferation Assay in the presence and absence of 50 µM of CH223191. <b>C.</b> The cells were grown in charcoaled stripped media and treated with DMSO (CON), 20 µM casodex (CDX), 50 µM CH223191 or a combination of CDX and CH223191 for 72 hours. Cell growth was measured using Promega CellTiter 96 Cell Proliferation Assay per manufactures instructions. Each bar represents mean±SEM (n = 3), *p<.05. <b>D.</b> Cells were serum starved for 24hrs and grown in charcoal stripped (CSS) media. Cells where then treated with DMSO (Con) or 50 µM CH223191 for an additional 24, 48 or 72 hours. At each endpoint, cells were analyzed for DNA content using the cell proliferation assay as described in the materials and methods. Each data point represents mean±SEM (n = 3). (*) denotes a significant difference compared to respective DMSO control.