Fig. 3

Hypoxic CM from GC cells induced NET formation through activation of the p38 MAPK pathway in neutrophils. A and B, Western blot analysis of p-ERK, p-p38, p-p65, p-Akt, and p-STAT3 levels in neutrophils cocultured with hypoxic-CM from GC cells for 2 h. C-E, Neutrophils were pretreated with inhibitors of the ERK, p38 MAPK, and p65 NF-кB pathways prior to incubation with hypoxic-CM from GC cells. The protein levels of p-ERK, p-p38, and p-p65 in neutrophils were measured by western blotting, and NET formation by neutrophils was evaluated with MPO and citH3 staining. Magnification: 20 × . Scale bars: 50 μm. All values are the means ± SDs. ns = not significant, *p < 0.05 and **P < 0.01 compared to the control group; #p < 0.05 and ##p < 0.01 compared to the hypoxic-CM (24 h) group