Figure 3: PUN induced HO-1 protein expression via PI3K/Akt pathway. Cells were incubated at 37°C in a humidified incubator with 5% CO2. (a) RAW264.7 cells were treated with 100 μM PUN for indicated durations (0, 2, 4, 6, 8, and 12 h). The Akt protein and Akt phosphorylation level were analyzed by Western blotting. (b) RAW264.7 cells were treated for 8 h with 100 μM PUN in the presence or absence of 20 μM LY294002. The HO-1 and Nrf2 proteins expression were analyzed by Western blotting. Data represent the mean ± SEM of three independent experiments and differences between mean values were assessed by one-way ANOVA. and indicate significant differences compared with the PUN-treated group of indicated proteins, respectively.