Oxidative Medicine and Cellular Longevity / 2013 / Article / Fig 5

Research Article

The NRF2 Activation and Antioxidative Response Are Not Impaired Overall during Hyperoxia-Induced Lung Epithelial Cell Death

Figure 5

The effects of acute and chronic hyperoxia on the binding of NRF2 to HMOX1 and NQO1 promoters. Cells were exposed to room air or hyperoxia for 1 to 12 h or for 36 to 48 h; chromatin was cross-linked and immunoprecipitated with IgG or anti-NRF2 antibodies, and DNA was amplified using gene specific primers. (a) Scheme showing the positions of the ARE sites, forward and reverse primers of HMOX1 and NQO1 promoters used in ChIP assays. (b) NRF2 binding to the HMOX1 and NQO1 promoters in cells exposed to acute hyperoxia (1–12 h). (c) NRF2 binding to the HMOX1 and NQO1 promoters in cells exposed to chronic hyperoxia (36 h or 48 h). PCR products were analyzed on 2% agarose gel. Band intensities were quantified with Image J software. Input DNA was used as a control. Graph represents mean ± SD of five independent experiments ( ). Fold increase was calculated over their respective room air controls , RA versus hyperoxia.

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