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. 2022 Mar 7;13:817596. doi: 10.3389/fphar.2022.817596

FIGURE 6.

FIGURE 6

DCL enhances the activation of Keap1/Nrf2/HO-1 in LPS/IFNγ-stimulated RAW264.7 macrophages. RAW264.7 cells were treated with DCL (1, 3, and 9 μM) or curcumin (5 μM) for 1 h, and followed by LPS/IFNγ incubation for additional 8 h. The protein expression of Keap1 (A), Nrf2 (C), and HO-1 (G) were measured by western blot. The densitometry analysis of Keap1 (B), Nrf2 (D), and HO-1 (H), normalized against GAPDH or α-Tubulin, respectively. The distribution of Nrf2 in cytoplasmic and nuclear fractions was analyzed using western blotting (E). The densitometry analysis of cytoplasmic and nuclear Nrf2, normalized against GAPDH and PARP1, respectively (F). (I,J) ROS production was measured using flow cytometry and relative statistical analysis. Data are represented as mean ± SEM, n = 3. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 compared to CK group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 compared to LPS/IFNγ group.