Fig. 4

Gastric cancer cell-derived exosomes induced pro-tumor activation of neutrophils through NF-κB pathway. a. Western blot assays for the expression of p-p65, p-STAT3, and p-ERK in neutrophils treated with GC-Ex for different times. b-h. Neutrophils were pre-treated with inhibitors for NF-κB, STAT3, and ERK pathways followed by incubation with BGC-Ex. The expression of phosphorylated NF-κB, STAT3, and ERK in neutrophils was determined by western blot (b). Western blot assays for the expression of LC3-II in neutrophils (c). The expression of ATG7 and BECN1 genes in neutrophils was determined by qRT-PCR (d). Flow cytometric analyses for apoptosis in neutrophils (e). The expression of CD11b in neutrophils (f). QRT-PCR analyses of pro-inflammatory factor gene expression (IL-1β, IL-6, IL-8, OSM, and TNFα) in neutrophils (g).Transwell migration assays for gastric cancer cells following treatment with supernatant from neutrophils (h). ^**P<0.01 and ^*P<0.05 compared to control (Ctrl); ^##P<0.01 and ^#P<0.05 compared to BGC-Ex