Fig. 2.
T-BHPL activated endothelial NADPH oxidase Endothelial cells were treated with t-BHP (50 μM) for 1 h with or without NAC (5 mM), Rot (20 μM), TTFA (10 μM), AA (5 μM), DPI (1 μM), ALL (10 μM) pretreatment 1 h. ROS production were evaluated by DCF (A and C) and apoptosis was detected by caspase 3/7 Activity Assay Kit (B). Cells were treated with t-BHPL for 1 h, the total, membrane and nucleus proteins were isolated. The expression of total NOX4 and p22 phox was detected by Western blotting (D and E). The location expression of NOX4 on membrane and in nucleus was detected with immunofluorescence (F) and Western blotting (G and H). Results were expressed as mean±SD of three independent experiments. AA, antimycin A; ALL, allopurinol; Cont, control group; DPI, diphenyleneiodonium; NAC, N-acetyl-L-cysteine; ROS, reactive oxygen species; Rot, rotenone; TTFA, 2-thenoyltrifluoroacetone.