BA6 Induces Apoptosis via Stimulation of Reactive Oxygen Species and Inhibition of Oxidative Phosphorylation in Human Lung Cancer Cells

Figure 5

Effect of BA6 induced on the mitochondria-apoptotic (intrinsic) pathway protein expression in A549 cells. (a) A549 lung cancer cells were treated with the indicated doses of BA6 for 24 h. However, cell lysates and protein were loaded to Western blot analysis using antibodies including Bax, Bcl-2, caspase-9, caspase-3, and β-actin antibodies. Here, β-actin was detected as the internal control. The Bcl-2 (b), Bax (c), procaspase-9 and cleaved caspase-9 (d), and procaspase-3 and cleaved caspase-3 (e) protein levels were quantified by ImageJ software, normalized with that of the β-actin level, and expressed as a normalization of the control. (f) The release level of cytochrome C from the mitochondria to the cytosol during BA6-induced apoptosis was performed by Western blot analysis. After the indicated treatment for 24 h, mitochondrial/cytosolic proteins were separated using mitochondrial/cytosol isolation kits and then subjected to Western blot analysis with antibodies, including COX IV, cytochrome C, and β-actin. (g) Protein levels of cytosolic and mitochondrial cytochrome C were quantified by ImageJ software, normalized with that of the β-actin and COX IV levels, and expressed as a normalization of the control. The data () are representative of three technical replicates. Asterisks indicate statistically significant differences between the treated group and the untreated group; and .

(a)

(b)

(c)

(d)

(e)

(f)

(g)