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. 2013 Jul 16;11(7):2574–2594. doi: 10.3390/md11072574

Figure 3.

Figure 3

APS8 induces apoptosis in NSCLC but not in normal fibroblasts. (A) Apoptosis after APS8 treatment (500 nM, 48 h) in A549, SKMES-1, and MRC-5 were assessed by staining with acridine orange and ethidium bromide and analysis by fluorescence microscope. Photos were taken at 400× magnification. Dashed arrows indicate cells in early apoptosis and full arrows point to late apoptotic cells. Green cells are alive; (B) Induction of apoptosis in A549, SKMES-1, and MRC-5 lines as measured by dual staining. Cells were treated with staurosporine (2 μM), APS8 (100 nM, 500 nM, and 1000 nM), nicotine (1 μM) or combination of APS8 and nicotine. The graph indicates the percentage of cells in the single cell populations. Each point is the mean of three independent experiments. The protective effect of nicotine was significant only for A549 cancer cells treated with 500 nM of APS8 (* P < 0.05); (C) APS8 induction of apoptosis in A549, SKMES-1, and MRC-5 cell lines was measured by flow cytometric analysis of annexin V and propidium iodide stained cells at 48 h. Controls (a, c, and e) and APS8 (500 nM) treated cells (b, d, and f); (D) Induction of apoptosis in A549, SKMES-1 and MRC-5 lines by flow cytometry. Cells were treated with staurosporine (2 μM), APS8 (100 nM, 500 nM, and 1000 nM), nicotine (1 μM) or combination of APS8 and nicotine. The graph indicates the percentage of gated cells in each cell population. Each point is the mean of three independent experiments.