Fig. 5.

SD2267 reduced APAP-induced ROS generation in hepatocytes

(A) AML12 cells were treated with 15 mM of APAP in the absence or presence of SD2267 for 24 h, and the cell viability was measured by MTT assay. Data are presented as means ± SEMs (n = 5 per group). (B) AML12 cells were treated with 5 mM of APAP in the absence or presence of SD2267, and then DCF-DA fluorescence intensities were measured at 4 h after APAP treatment. (C) AML12 cells were treated with 5 mM of APAP in the absence or presence of SD2267 for 8 h, and then stained with MitoSOX Red. The intensities of MitoSOX Red were measured by flow cytometry, and presented as means ± SEMs (n = 3 per group). (D) AML12 cells were treated with 5 mM of APAP in the absence or presence of SD2267 for 8 h, and then stained with JC-1. The green fluorescence intensities of JC-1 were measured by flow cytometry, and presented as means ± SEMs (n = 3 per group). (E) Primary mouse hepatocytes (PMHs) were treated with SD2267 at various concentrations for 24 h, and KEAP1 expressions were determined by western blotting. Densitometry was performed using three independent experiments and presented as a graph with DC₅₀ and D_max value. (F) PMHs were treated with 100 nM of SD2267 for 6 h before APAP treatment at 20 mM, and then DCF-DA fluorescence intensities were measured every 20 min for 10 h. Data are presented as means ± SEMs (n = 4 per group). *p < 0.05, **p < 0.01, ***p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)