FIG. 9.
The effects of Nrf2 knockdown on MeHg-induced Ho-1, Nqo1, and xCT expression were analyzed by real-time PCR. Primary microglial cells were first infected with lentivirus as indicated for 24 h and then treated with 5μM MeHg for 30 min. Then the mRNA levels of Ho-1 (A), Nqo1 (B), and xCT (C) were measured by real-time PCR. The difference in the average threshold cycle (ΔCt) values was determined and normalized to the expression of β-actin. The uninfected and untreated microglial cells were used to determine the basal gene expression levels. The uninfected cells treated with 5μM MeHg for 30 min served as a positive control. The random virus with no known gene target was also used to infect microglial cells in order to reveal any possible effects of lentiviral backbone on gene expression. Microglial cells treated with Nrf2-specific shRNA showed no significant increase in downstream gene expression compared with basal levels after 5μM MeHg treatment for 30 min. Values are expressed as the mean ± SEM derived from three independent experiments.