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. 2019 Jan 30;54(4):1271–1281. doi: 10.3892/ijo.2019.4704

Figure 8.

Figure 8

Effect(s) of metformin on mitochondria-mediated caspase-dependent apoptotic signaling of AGS cells. The cells were treated with 0, 10, 20 and 30 mM of metformin for 48 h, and whole-cell lysates and mitochondrial and cytosolic fractions were then harvested. Protein levels of (A) p-BAD, BAD and Bcl-2, (B) Apaf-1 and (C) caspase-9, caspase-3 and caspase-7 signals were determined by western blot analysis. β-actin was an internal loading control. (D) The cytosolic (top) and mitochondrial (bottom) fractions were used to determine for cytochrome c translocation by western blot analysis. GAPDH and COX IV were internal loading controls. (E) Following pre-incubation with or without 10 µM z-VAD-fmk (a pan-caspase inhibitor) for 2 h, the cells were exposed to 30 mM metformin for 48 h. Cell viability was assessed using the MTT assay. The values are presented as the mean ± standard deviation of triplicates. *P<0.05 vs. metformin-treated only. p-, phospho; BAD, Bcl-2-associated agonist of cell death; Bcl-2, B-cell lymphoma-2; Apaf-1, apoptotic protease-activating factor-1; z-VAD-fmk, carbobenzoxyvalyl-alanyl-aspartyl fluoromethyl ketone.