BioProject

The overall cellular response to oxidative stress generated by Ero1α in the lumen of the mammalian endoplasmic reticulum (ER) is poorly characterized. More...

The overall cellular response to oxidative stress generated by Ero1α in the lumen of the mammalian endoplasmic reticulum (ER) is poorly characterized. Here, we investigate the effects of overexpressing a hyperactive mutant (C104A/C131A) of Ero1α. Using microarray analysis, we demonstrate that the cell reacts to the oxidative challenge caused by Ero1α hyperactivity by turning on the unfolded protein response. Our findings suggest that the hyperoxidation generated by Ero1α-C104A/C131A is addressed in the ER lumen and is unlikely to exert oxidative injury throughout the cell. Overall design: Human embryonic kidney 293 Flp-In T-REx cells (Invitrogen) overexpressing Ero1α-WT and Ero1α-C104A/C131A were grown in triplicates and expression was either not induced or induced doxycycline (dox) for 24 h. Extracted RNA was hybridized on Affymetrix microarrays and genes with significantly different expression levels between the four categories were identified by a two-way analysis of variance (ANOVA), where the cell line (Ero1α-WT or Ero1α-C104A/C131A) and induction status (-dox/+dox) were used as the two factors for the p-value calculation. Less...