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. 2018 Sep 6;19(9):2649. doi: 10.3390/ijms19092649

Figure 2.

Figure 2

(A) The dose effect of lakoochin A at 24 h on the mitochondrial membrane potential (∆Ψm) of A375.S2 cells, as determined by flow cytometry staining with JC-1. (B) The time effects of lakoochin A on the ∆Ψm of A375.S2 cells pre-labeled with 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanineiodide (JC-1) (10 μg/mL) for the indicated times (0.5–16 h). (C) Effect of lakoochin A on mitochondrial reactive oxygen species (ROS) production (determined by flow cytometry after staining with MitoSOX Red indicator) in A375.S2 cells. (D) Effect of lakoochin A on cellular ROS production (determined by flow cytometry after staining with H2DCFDA reagent). (E) The cellular ROS production of several antioxidants and lakoochin A, determined by flow cytometry staining with H2DCFDA reagent. The A375.S2 cells were pretreated for 1 h with mitochondria-targeted antioxidant (MitoTEMPOL), antioxidant (NAC), or nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (DPI) and then treated with lakoochin A for 4 h. (F) Effect of MitoTEMPOL, NAC, and DPI on lakoochin A-induced A375.S2 cell apoptosis (determined by MTT assay). The data were collected from at least three individual experiments and expressed as mean ± S.E.M. * p < 0.05, # p < 0.01 compared to the control group.