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Figure 5: Blocking Nrf2 suppresses modulatory effect of PI3K/Akt pathway on PUN-induced HO-1 protein expression. RAW264.7 cells were treated for 8 h with 100 μM PUN in the presence or absence of indicated inhibitor (LY294002, 20 μM; brusatol, 10 nM) and inducer (insulin, 10 μM) at 37°C in a humidified incubator with 5% CO2. The HO-1 protein expression was analyzed by Western blotting. Data represent the mean ± SEM of three independent experiments and differences between mean values were assessed by one-way ANOVA. indicates significant differences compared with the PUN-treated group.