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. 2017 Jan 5;2017:3692797. doi: 10.1155/2017/3692797

Figure 3.

Figure 3

Combination treatment with BZYQD and cisplatin leads to apoptosis induction in A549/DDP cells. (a) A549/DDP cells were pretreated with 100, 250, and 500 μg/ml BZYQD for 2 h, and then 40 μg/ml cisplatin was added for another 24 h. Photomicrographs of representative fields of cells stained with Hoescht-33258 to evaluate nuclear chromatin condensation. (b) The percentage of AnnexinV+/PI− (early apoptotic cells, lower right), AnnexinV+/PI+ (late apoptotic cells, upper right), AnnexinV−/PI− (viable cells, lower left), and AnnexinV−/PI+ (necrotic cells, upper left) cells is shown. (c) Cell lysates were prepared and subjected to immunoblotting with antibodies to cleaved caspase 3, cleaved-PARP, Bcl-2, Bax, and GAPDH. (d) A549/DDP cells were pretreated with 20 μM z-VAD-fmk 2 h prior to cotreatment with BZYQD and cisplatin. After incubation, cell growth inhibition was determined by the Cell Counting Kit as described previously. The results are expressed as the mean ± SD of four independent experiments. (ANOVA followed by Bonferroni's test.) BZYQD: Bu-Zhong-Yi-Qi decoction; PI: propidium iodide; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; z-VAD-fmk: z-Val-Ala-Asp-fluoromethylketone.