The NRF2 Activation and Antioxidative Response Are Not Impaired Overall during Hyperoxia-Induced Lung Epithelial Cell Death
Figure 2
NRF2 target gene expression in AECs exposed to acute hyperoxia. Cells were exposed to either room air or hyperoxia for 1 to 12 h and then harvested at indicated time points for total RNA, and protein extraction. (a) cDNA was prepared from total RNA and the mRNA expression levels of HMOX1, GCLC and NQO1 were quantified using TaqMan assay probes. The values represent the mean ± SD of three independent experiments (). RA versus hyperoxia. (b) A comparable amount of whole cell extracts (~40 μg) was separated on SDS-PAGE gel and immunoblotted with anti-HMOX1, anti-NQO1, or anti-GCLC antibodies. -actin was used a as reference. Graph represents the relative band intensities of the HMOX1, NQO1, and GCLC of three independent experiments done in duplicate (). The band intensities were quantified using Image J software and normalized with their respective -actin band intensities. Band intensities of room air exposed samples were considered as one arbitrary unit. , RA versus hyperoxia.
