Figure 3

Biliverdin regulates NR2E3 in cells. (A, B) Reporter gene expression analysis in 293F cells. Cells were transfected with a reporter plasmid encoding nanoluciferase under the control of 9 × GAL^UAS and plasmids (pFN26A derivatives) encoding GAL^DBD-NR2E3 fusion protein. A day after transfection, cells were treated with vehicle (0.1% DMSO) or biliverdin for 3 h. Nanoluciferase activity was measured using Vivazine substrate (average and standard deviations, n = 3). (C, D) Biliverdin increased NR2E3 protein level in WERI-Rb-1 cells. WERI-Rb-1 human retinoblastoma cells were incubated with vehicle (0.1% DMSO) or biliverdin (1 μM for C and varying concentrations for D) for 24 h. Western blot results (1:1000-fold diluted antibody) are shown. COX IV was used as a loading control. In Fig. 3C, different parts of the same gel are shown. Full-sized Western blot images are available in the Supplementary Information.