FIG. 6.

Effects of Nrf-2 transfection on expression of Nrf-2 and its downstream target genes in HEK293 cells and C1 astrocytes. (A) HEK293 cells were either mock transfected with the empty vector pcDNA3.1 or transfected with the pCMV-Nrf-2 vector and then incubated for 48 h prior to preparation of cell lysates for analysis of their Nrf-2, xCT, γ-GCLC, γ-GCLM, and GPx content. (B) C1 cells were either mock transfected with the empty vector pcDNA3.1 or transfected with the pCMV-Nrf-2 vector and then cultured for 56 h either without further treatment (CON), or with BSO or NAC, as indicated in Materials and Methods. At the end of the culture period, total cellular lysates were analyzed for their Nrf-2, xCT, γ-GCLC, γ-GCLM, and GPx content. All blots were stripped and reimmunoblotted with anti-β-actin antibody as a protein loading control. Intensities of bands corresponding to proteins of interest were normalized to those for β-actin, and differences determined as shown. All blots shown are representative of three independent experiments.