Journal of Cancer Prevention : pISSN 2288-3649 / eISSN 2288-3657

Fig. 2.

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Fig. 2. Nuclear translocation and transcriptional activity of nuclear factor E2-related factor-2 (Nrf2) and its implication in NAD(P)H:quinone oxidoreductase-1 (NQO1) upregulation induced by Z-ajoene. (A) Nuclear extracts and cytosol fraction from cells treated with Z-ajoene at indicated concentrations for 20 hours were subjected to Western blot (WB) analysis. Data are representative of three independent experiments showing a similar trend. (B) MCF-10A cells transiently transfected with nqo1-luciferase construct harboring antioxidant response element (ARE) sequences or mock vector were incubated with Z-ajoene (10 or 20 μM) for 20 hours as described in Materials and Methods. Transcriptional activity of nqo1-specific ARE was measured by the luciferase reporter gene assay with a luminometer. Error bars represent means ± SD of three independent experiments. Asterisks indicate the statistically significant difference as compared with control (*P < 0.05; **P < 0.01). (C) MCF-10A cells transiently transfected with Nrf2-siRNA or control siRNA were incubated with or without 20 μM Z-ajoene for 20 hours. Cell lysates were separated by SDS-PAGE (for Nrf2, 8%; for NQO1, 12%) and immunoblotted with NQO1 and Nrf2 antibodies. Actin was served as a loading control. ***Significantly different from the untreated control (P < 0.001). (D) MEF cells (wild type and Nrf2−/−) were treated with or without 20 μM Z-ajoene for 20 hours. Cell lysates were conducted immunoblotting. The band intensity of immunoblots for NQO1 was normalized to that of actin by GelPro image densitometry followed by statistical analysis. Error bars represent means ± SD of three independent experiments. ***Significantly different from the untreated control (P < 0.001).
Journal of Cancer Prevention 2019;24:112-22 https://doi.org/10.15430/JCP.2019.24.2.112
© 2019 Journal of Cancer Prevention