Fig. 4. Effect of GA or CDDP on apoptosis of A549 cells. (A) A549 cells were seeded at a density of 8×104 cells per well in cell culture dishes and treated with NC (0.1% DMSO), CDDP (12 μM) and GA (75 μM) for 48 h, and subsequently subjected to Annexin V assay. The cell distribution of apoptosis was analyzed and quantified. (B) A549 cells were seeded at a density of 3×104 cells per well in 24-well plates and treated with NC (0.1% DMSO), CDDP (12 μM) and GA (75 μM) for 48 h; TUNEL assay was conducted and cells were observed under ×100 magnification. DNA fragmentations were stained with TUNEL and all nuclei are stained with DAPI. DNA fragmentations in nuclei were shown in MERGE. The percentage of TUNEL-stained cells for DAPI-stained cells was analyzed using Image J program. A549 cells were seeded at a density of 6×105 cells in cell culture dishes, and treated with NC (0.1% DMSO), CDDP (12 μM) and GA (75 μM) for 48 h. (C) The expressions of apoptosis related proteins such as p53, Bax, Bcl-2, and cleaved caspase-3 were determined by Western blot assay. (D) The expression levels of each protein were normalized to GAPDH and quantified. The representative images were obtained from at least three repeated experiments. Data are expressed as means ± SD from three independent experiments. *p<0.05 vs. NC (0.1% DMSO).
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