Glis mediate Hh-induced transactivation of P450scc, 3β-HSD1 and aromatase in JEG-3 cells. (A–C) Dual-luciferase assays in JEG-3 cells, after infection with Gli-shRNA-expressing lentivirus, transient transfection with P450scc, 3β-HSD1 or aromatase reporter constructs, and treatment with either control medium (−) or Shh conditional medium (SM, +) for 48 hrs. (D–F) Dual-luciferase assays in JEG-3 cells, 48 hrs after transient co-transfection with constitutively active forms of Gli (Gli3-ΔC, ΔN-Gli2) and reporter constructs of either wild type or Gli binding site mutants for P450scc, 3β-HSD1 and aromatase. (G–I) Detection of the interactions between Glis and DNA of P450scc, 3β-HSD1 and aromatase by ChIP assays in JEG-3 cells, after 48 hrs of treatment with either control medium (CM) or Shh conditional medium (SM). (J) A model for Hh-induced steroidogenesis in human trophoblasts. **, ‡p<0.01, *,†
p<0.05; n=6, error bar, SD.