FIG. 5.

Activation and nuclear accumulation of Nrf-2 in ts1-infected or cystine-deficient astrocytes. (A) After mock, WT, or ts1 infection, C1 cells were incubated for 24, 48, and 72 hpi, and total cell lysates analyzed for their contents of Nrf-2. (B) Mock- or ts1-infected C1 cells were incubated for 24, 48, and 72 hpi, at which time cell homogenates were separated into cytoplasmic and nuclear fractions. The separate fractions were then analyzed for their contents of cytoplasmic and nuclear Nrf-2. (C) Mock-, WT-, or ts1-infected C1 cells were incubated for 72 hpi, and their nuclear fractions were analyzed for their contents of Nrf-2. (D) PACs were mock or ts1 infected either in the presence or absence of BSO and incubated for 72 hpi. Cytosolic and nuclear fractions were prepared and analyzed for their contents of cytosolic and nuclear Nrf-2. (E) Uninfected C1 cells incubated for 24 h in medium containing the indicated concentrations of cystine were analyzed for their total cellular contents of Nrf-2. 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 to four independent experiments.