Fig. 4

Gastrin induced survival is dependent on autophagy. (a) AGS-Gr cells treated with gastrin +/− BafA1 for 18 h. Cell viability was assessed using annexin V-PI staining and flow cytometric analyses. The viability of untreated cells (U.S.) is set to 1.0. (b) Gastrin reduces cisplatin induced cell death. AGS-Gr cells treated with increasing doses of cisplatin (7.5-90 μM) in presence or absence of gastrin (10 nM). Cell viability assessed by XTT assay; the viability of cisplatin treated cells is set to 1.0 for each concentration (c) AGS-Gr cells treated with gastrin (2 h) with subsequently treatment with cisplatin (7.5 μM) for 36 h. Autophagy was blocked for 12 h using HCQ. Cell viability was determined by XTT assay; the viability of untreated cells (U.S.) set to 1.0. (d) Cells treated with gastrin for (2 h) and subsequently with increasing concentrations of cisplatin (1–7.5 μM) for 36 h before autophagy was blocked for 12 h. (e) Gastrin induced survival is dependent on ULK1. AGS-Gr cells treated with ULK1 inhibitor SBI-026965 (SBI) (5 μM) for 24 h and 48 h in the presence or absence of gastrin (10 nM). (f) Caspase activity performed with AGS-Gr cells pretreated with gastrin (2 h), followed by cisplatin (10 μM) treatment for 72 h. Autophagy was blocked for 12 h with BafA1. Bar graphs (a, b c, d, e and f) represent SEM (n = 3, P-value: * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001)