SSE induces both apoptotic and autophagic cell death. (A) AGS cells were treated with SSE (25 and 50 μg/mL) for 6, 12, and 24 h, stained with YO-PRO-1, and analyzed using flow cytometry. Histogram and density plot data are shown. (B) For the detection of chromatin condensation, cells treated with SSE (50 μg/mL) for 24 h were stained with DAPI and observed under a fluorescent microscope. (C) Cells seeded on cover slips were transiently transfected with RFP-tagged LC3 plasmid DNA (RFP-LC3), treated with SSE (50 μg/m) for 12 h, and observed for fluorescence. (D) Caspase-3 activation, PARP cleavage, increases in Beclin-1, and LC3-II expression were assessed using western blot analysis after treatment with SSE (25 and 50 μg/mL) for 12 and 24 h. Changes in Bcl-2 and Bax expression after SSE treatment were also determined. Data represent 3 independent experiments.