Figure 5
Differential regulation of Slc7a11 promoter activity in WT and Jdp2-KO GCPs by Jdp2, Nrf2, and p21Cip1 through the ARE. (a) Schematic representation of the mouse Slc7a11 promoter and the positions of qPCR primers for the ChIP assay. ARE, antioxidative responsive element, NS, non-specific sequence motifs (as a negative control). (b) Slc7a11 promoter–luciferase activity (pGL3-4.7); mouse xCT–luciferase (ref. 10) was elevated in Jdp2-KO GCPs. Each value represents the mean ± SEM (n = 3); *p < 0.05. (c) ChIP–qPCR analyses were performed on chromatin extracts from WT and Jdp2-KO GCPs using the indicated antibodies and normal IgG (as a negative control) as described in Materials and Methods. Each value represents the mean ± SEM (n = 5); *p < 0.05. (d) and (e) Effects of Jdp2 and p21Cip1 on ARE–luciferase activity. pGL-hQR25-luciferase (100 ng) plus 0–400 ng of pcDNA-Jdp2 (d) or pcDNA-p21Cip1 (e) were transfected into Jdp2-KO GCPs. One day after transfection, cells were collected, and luciferase activity was measured as described in Materials and Methods. Values represent the mean ± SEM. (n = 3). *p < 0.05.
