Figure 2.

FasL⁺PD‐L2⁺ neutrophil subset with a unique phenotype is increased in GC as tumor progresses and predicts poor patient survival. A) Statistics analysis of FasL⁺PD‐L2⁺ neutrophil percentage in total neutrophils or FasL⁺PD‐L2⁺ neutrophil number per million total cells in each samples of patients with GC (n = 51). B) Dot plots of surface molecule staining for FasL⁺PD‐L2⁺ neutrophils gating on CD45⁺CD11b⁺CD66b⁺CD15⁺ cells. C) Representative image of MPO⁺FasL⁺ neutrophils and MPO⁺PD‐L2⁺ neutrophils in tumor tissues of GC patients by immunohistochemical staining. D) FasL⁺PD‐L2⁺ neutrophil percentage and FasL⁺PD‐L2⁺ neutrophil number among TNM stages was compared. Kaplan‐Meier plots for overall survival by median FasL⁺PD‐L2⁺ neutrophil percentage (36.8) and FasL⁺PD‐L2⁺ neutrophil number (306). E) PPI network analysis of significantly changed genes from top 15 gene ontology terms in tumor‐infiltrating FasL⁺PD‐L2⁺ neutrophils, as compared with peripheral FasL⁻PD‐L2⁻ neutrophils. F) Gene ontology analysis of significantly changed genes in tumor‐infiltrating FasL⁺PD‐L2⁺ neutrophils, as compared with peripheral FasL⁻PD‐L2⁻ neutrophils. G) Differentially expressed genes between tumor‐infiltrating FasL⁺PD‐L2⁺ neutrophils and peripheral FasL⁻PD‐L2⁻ neutrophils are shown. H) Heatmap revealing gene changes between tumor‐infiltrating FasL⁺PD‐L2⁺ neutrophils and peripheral FasL⁻PD‐L2⁻ neutrophils from five GC patients. Data are mean ± SEM and analyzed by Student's t‐test, Mann‐Whitney U‐test, and one‐way ANOVA. *P < 0.05, **P < 0.01 for groups connected by horizontal lines. MPO, myeloperoxidase; FasL⁺PD‐L2⁺ Neu (%), FasL⁺PD‐L2⁺ neutrophil percentage; FasL⁺PD‐L2⁺ Neu (NO), FasL⁺PD‐L2⁺ neutrophil number.