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Figure 1: PUN increases Nrf2/HO-1 expression in RAW264.7 cells in time-dependent manner and dose-dependent manner. 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, 12, and 24 h). The HO-1 protein expression was analyzed by Western blotting. (b) RAW264.7 cells were treated for 8 h with PUN at the indicated concerntrations (0, 5, 25, 50, 100, and 200 μM). The HO-1 protein expression was analyzed by Western blotting. (c) RAW264.7 cells were treated with 100 μM PUN for indicated durations (0, 2, 4, 6, 8, 12, and 24 h). The Nrf2 and Keap1 protein expression were analyzed by Western blotting. (d) RAW264.7 cells were treated for 8 h with PUN at the indicated concerntrations (0, 5, 25, 50, 100, and 200 μM). The Nrf2 and Keap1 protein 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 control group of indicated proteins, respectively.