Fig. 3. The Srxn1 promoter contains an ARE which is induced by Nrf2 expression in cortical neurons, as is the endogenous gene.
A) Schematic showing the putative ARE within mammalian Srxn1 promoters. B) Effect of Nrf2 expression on wild-type and mutant Srxn1-Luc reporters (normalized to Renilla control-see methods). Neurons were transfected with Srxn1 reporter plus pcDNA3.1-Nrf2 or pGlobin control plasmid. NB. In this and all experiments “con” denotes pGlobin control vector. *p<0.05 (2-tailed paired T-test, n=3-5). C) Effect of Nrf2 expression on ARE-Luc activity. *p<0.05 (n=5). D) Effect of transfecting pEF-Nrf2 on endogenous Srxn1 expression. Neurons were transfected with the indicated vectors and after 24 h subjected to immunocytochemical analysis of sulfiredoxin expression. Immunofluorescence performed as described (McKenzie et al. 2006). Scale bar = 40 μm. E) Effect of Nrf2 expression on WT and mutant Srxn1-Luc reporters in glial cells *p<0.05 (n=7). F) Effect of Nrf2 expression on ARE-Luc activity in glial cells. *p<0.05 (n=4). G) Effect of Nrf2 expression on WT and mutant Srxn1-Luc reporters in HEK293cells. *p<0.05 (n=3).