Figure 3: nsPEFs and everolimus induced melanoma cell apoptosis, cellular mitochondrial membrane potential (ΔΨm) loss and Ca2+ increase.
(a) A375 cells were treated with everolimus for 48 h, followed by nsPEF treatment. Cell apoptosis was evaluated by FITC-Annexin V and PI staining using flow cytometry. The experiments repeated 3 times yielded similar results. Cells after nsPEF treatment were incubated with JC-1 or Fluo-3/AM at 37 °C for 30 min. (b) Loss of ΔΨm was detected using the fluorescent probe JC-1. Cells treated with 100 nM valinomycin were used as the positive control for decreased ΔΨm. (c) Intracellular Ca2+ level was quantified with the fluorescent dye Fluo-3/AM. Cells treated with 20 μM ionomycin were used as the positive control for intracellular Ca2+ level detection. The experiments repeated thrice yielded similar results. Values are means ± SD (n = 6). *P < 0.05 compared to control, #P < 0.05 compared to everolimus single agent treatment.
