Nur77 increases relaxin-3 expression at transcriptional levels.
A, localization of the NBRE in the promoter region of human, rat, and mouse RLN3; B, schematic representation of relaxin-3 promoter and its mutants. C, Nur77 increases RLN3 promoter activity in a dose-dependent manner. NRVMs cultured in 12-well plates were transiently transfected with 200 ng of RLN3 promoter luciferase reporter vector and 20 ng of pCMV-R.Luc, together with the indicated amount of Nur77 expression vector. 48 h after transfection, NRVMs were washed once with ice-cold PBS and then analyzed for luciferase reporter activities using the dual luciferase reporter assay system (Promega) (n = 4). *, p < 0.05 compared with Ad-LacZ group. D, Nur77 binds to the RLN3 promoter. NRVMs were transduced with the indicated adenoviruses at an MOI of 50. 48 h after transduction, the recruitment of Nur77 to the RLN3 promoter was determined by ChIP assays. Error bars, S.D.