FIGURE 3.

Cul3 and Rbx1 WT AREs and not mutant AREs are inducible by antioxidants. A, ARE-luciferase expression in transfected cells. ARE-5 for both Cul3 and Rbx1 were separately attached to SV40 basal promoter hooked to luciferase reporter gene by cloning in vector pGL2 promoter, wild type and mutant sequences are shown. B and C, wild type and mutant Cul3 and Rbx1 AREs were transfected in Hepa-1 cells, treated with DMSO, or t-BHQ (100 μm for 16 h), and analyzed for luciferase activity. For all the above experiments, pGL2 empty vector was used as negative control. The results are expressed as fold increase in relative luciferase activity compared with untreated pGL2 transfection. The data shown are mean ± S.D. of three independent transfection experiments in A–C.