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. 2014 Dec 1;36(1):131–138. doi: 10.1038/aps.2014.108

Figure 5.

Figure 5

DMAS-induced A549 cell apoptosis is mediated through p38 activation. (A) A549 cells were pretreated with or without SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measured as controls. (B) A549 cells were pretreated or not with SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (C) A549 cells were pretreated or not with SP600125 (25 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (D) A549 cells were treated with DMAS in the presence or absence of SB203580 (10 μmol/L) for 24 h. The apoptotic status was determined using an Annexin V-FITC binding assay. (E) A549 cells were pretreated with or without Z-VAD-FMK (10 μmol/L), followed by treatment with DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measure as controls.