Fig 4.

Genipin increases oxaliplatin-induced cell death via autophagy. (A) AGS cells were treated with genipin 100 µM for 24h. The cells were observed by light microscopy. Scale bar: 20 μm. (B) The autophagy was observed by immunofluorescence using autophagy detection kit (original magnification: 40×). Scale bar: 10 µM. (C) AGS cells were treated with genipin 100 µM for 24h. The protein expression of Beclin1, p62, LC3, AMPK β1, AMPK β2, and DRAM were measured by western blotting. β-Actin was used as a loading control for each lane. (D) The LC3 puncta were observed by immunofluorescence (original magnification: 40×). Scale bar: 10 µM. (E) AGS cells were transfected with control siRNA or LC3 siRNA and then the cells were treated with oxaliplatin, genipin, or combination. The activity of cleaved-PARP and cleaved-caspase 3, and cleaved-caspase 9 were measured by western blotting. (F) AGS cells were transfected with control siRNA or LC3 siRNA and then the cells were treated with oxaliplatin, genipin, or combination. The cells were stained with annexin V and PI and then were measured using FACS analysis. (G) Schematic diagram for combination model of oxaliplatin and genipin. ***P < 0.001, *P < 0.05.