Figure 2. LPS induces de novo synthesis of Nrf2.
THP-1 cells were stimulated with LPS (10 μg/ml) for various times and whole cell extracts prepared and separated by SDS-PAGE for Western blot analysis of A-C Nrf2 or D Keap-1 protein expression. THP-1 cells were pre-treated with either B cycloheximide (CHX)) or C actinomycin D (Act D) respectively for 1 h before LPS treatment and western blot analysis of Nrf2 protein expression. Chemiluminescent images were quantified by densitometry. The numerical values represent fold increase in expression following stimulation with LPS compared to no LPS, and normalized to β-actin as a loading control. E, THP-1 cells were stimulated with LPS for various times and RNA extracts prepared. Nrf2 mRNA expression was measured by real-time PCR. Data shown represents results from three experiments (mean ± SD).