Protective Role of Nuclear Factor E2-Related Factor 2 against Acute Oxidative Stress-Induced Pancreatic β-Cell Damage
Stable knockdown of Nrf2 results in reduced expression of ARE-dependent genes, decreased intracellular GSH, and elevated intracellular ROS levels in MIN6 cells. (a) mRNA expression of Nrf2 in cells transduced with lentiviral shRNA targeted against mouse Nrf2 (Nrf2-KD) or scrambled nontarget negative control (Scr). (b) Reduced protein expression of NRF2 in Nrf2-KD cells under basal and arsenite (5 μM), tert-butylhydroquinone (tBHQ, 50 μM) treated conditions. Cells were exposed to the chemicals for 6 hrs. Whole cell lysates (WL) and nuclear protein (NP) were used for immunoblotting, and β-Actin and Lamin A were used as loading controls, respectively. (c) Expression of ARE-dependent genes under vehicle (Veh) and tBHQ treated conditions. Cells were treated with the tBHQ for 6 hrs. The expression of Gclc and Nqo1 was measured by real-time RT-PCR. (d) Intracellular GSH levels. (e) Intracellular ROS levels. Values in (a), (c), (d), and (e) are means ± SD. . versus Scr with the same treatment.
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