AKR1B10 was upregulated via NRF2 upon oxidative stress. A. ROS assay in Huh7 cells with Dox (0.5 μg/mL) treatment for 36 hours. DCFH-DA was the probe targeting the ROS. The scale bar is 200 μm. B, C. RT-qPCR and western blot assays of AKR1B10 in Huh7 cells after treatment with Dox (0.5 μg/mL) for 16 hours and 36 hours, respectively. D. Schematic of NRF2 transcriptional regulation element and the site-directed ARE mutation located in AKR1B10 promoter. The nucleotides in red indicated the mutant sites compared to the wild type AKR1B10 promoter sequence. E. Luciferase activity assay detected in Huh7 cells with AKR1B10 promoter and NRF2 expression plasmids transfection. F. Western blot assay of AKR1B10 protein level with NRF2 overexpression. Data showed the mean ± s.d. of three independent experiments (*P<0.05, **P<0.01, Student’s t-test, two-tailed).