Fig. 5: The PI3K/AKT pathway is involved in chaetocin-induced apoptosis in GC cells.

a RNA-seq was performed on non-treated AGS cells as well as AGS cells with 100 nM chaetocin treatment for 12 h. And DEGs defined as +/− 2-fold change and FDR < 0.01 were included for enriched gene pathway analysis. b p-AKT and AKT expression levels were analyzed by western blot in chaetocin-treated HGC-27 and AGS cells. c AKT and flag-tag expression levels were detected by western blot in HGC-27 and AGS cells transiently overexpressing AKT. AKT-overexpressing HGC-27 and AGS cells were treated with chaetocin (100 nM for HGC-27, 50 nM for AGS). d Apoptosis was analyzed by flow cytometry. Results were shown as mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. e Cleaved-PARP and caspase-3 expression levels were determined by western blot. HGC-27 and AGS cells were pretreated with 20 μM LY294002 for 2 h and then cotreated with chaetocin (100 nM for HGC-27, 50 nM for AGS). (f) Apoptosis was analyzed by flow cytometry. Results were shown as mean ± SD of three independent experiments. ***P < 0.001. g Cleaved-PARP and caspase-3 expression levels were determined by western blot. (h) HGC-27 and AGS cells were pretreated with 5 mM NAC for 1 h and then cotreated with chaetocin (200 nM for HGC-27, 100 nM for AGS). Then, p-AKT expression levels were analyzed by western blot. All blots presented here are representative of three independent experiments