Nrf2 expression induces ALR expression. (A) For reporter-gene assay HepG2 and Huh-7 cells were cotransfected with luciferase-reporter construct harboring ALR promoter (−733 to +240) with putative ARE sequence and caNrf2 or dnNrf2 expression plasmids (all 0.5 µg each). Cotransfection with pcDNA3 vector served as control and was set 1 (three independent experiments in triplicates, mean ± SEM). (B) For mRNA expression analysis, HepG2 and Huh-7 cells were transfected with caNrf2 or dnNrf2 expression plasmid (1 µg, 24 h) and mRNA expression of ALR as well as NQO1 were analyzed by quantitative RT-PCR. Transfection with pcDNA3 vector served as control and was set 1. 18S ribosomal RNA expression was determined for normalization (two independent experiments in triplicates, mean ± SEM). (C) Western blot analysis of cellular lysates of HepG2 cells transfected with caNrf2 or dnNrf2 expression plasmids was performed followed by densitometric analysis of 21- and 23-kDa ALR band. *P < 0.05 differs from corresponding control.