Figure 6

Downregulation of Nrf2 impairs protection of astrocytes against iron overload. (A) The expression of Nrf2 was analyzed by RT-qPCR in resting (C) and cytokine-activated (CK) astrocytes. Statistical analysis was calculated by using paired two-tailed Student’s t-test. (*p < 0.05). (B) Resting astrocytes were incubated with lipofectamine alone or transfected with a mix of two Nrf2-specific siRNAs for 90 min. After at least 6 h from transfection, astrocytes were treated with proinflammatory cytokines to induce activation. Twenty-four hours later, an acute iron overload protocol was performed, and after 90 min cell viability (MTT assay) was measured on resting (C) or activated astrocytes (CK) subjected or not to Nrf2 silencing. Statistical analysis was performed by using one-way ANOVA followed by Bonferroni post hoc test. (*p < 0.05; **p < 0.01; ***p < 0.001).