NR4A1 plays a role in RCC proliferation.

<p>(A) Pathways activated by NR4A1 and targeted by NR4A1 antagonists. (B) Cells were transfected with two different oligonucleotides targeting NR4A1 [NR4A1(1) and NR4A1(2)] and after 72 hr, the number of cells were determined as outlined in the Materials and Methods. (C) Cells were treated with DIM-C-pPhOH and DIM-C-pPhCO₂Me and cell numbers were determined after treatment for 24 hr. (D) Cells were transfected with NBRE₃-luc and 40 ng FLAG-NR4A1, treated with DMSO, DIM-C-pPhOH (20 μM) and DIM-C-pPhCO₂Me (15 μM), and luciferase activity was determined as described [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128308#pone.0128308.ref017" target="_blank">17</a>]. (E) Cells were transfected with siCtl (non-specific) or siNR4A1 and treated with DIM-C-pPhOH and DIM-C-pPhCO₂Me and cell numbers were determined. siNR4A1 alone decreased cell proliferation as indicated in (B) and this value was set at 100% to determine the effects of C-DIMs in cells after loss of NR4A1. (F) Athymic nude mice bearing ACHN cells as xenografts were treated with 30 mg/kg/d DIM-C-pPhOH and tumor volumes were determined. Results (C—E) are means ± SE for at least 3 separate determinations and significantly (p < 0.05) decreased growth/volume is indicated (*). Significant (p < 0.05) attenuation of C-DIM-induced growth or luciferase activity or after transfection with siNR4A1 is also indicated (**).