Figure 3

Astrocytes prolong neutrophil survival in a cell-cell contact-dependent manner. A & C. FACS-sorted resting or LPS-primed neutrophils were co-cultured with resting or poly I:C-primed astrocytes at a ratio of 1:1 for 16 hr. After co-culture, all cells were trypsinized, washed with PBS once and the neutrophils were immunoreacted with APC-conjugated anti-mouse CD11b antibody. CD11b⁺ Neutrophils were then stained with Annexin V and propidium iodide (PI) for detection of apoptosis and necrosis. White bar, neutrophils alone; gray bar, neutrophils co-cultured with resting astrocytes; black bar, neutrophils co-cultured with primed astrocytes (*p < 0.05, vs resting neutrophils alone; **p < 0.01, vs resting neutrophils alone; ++p < 0.01, vs primed neutrophils alone. n = 5/bar). B & D. Resting neutrophils were cultured alone or cultured with astrocytes in the same (co-culture) or separate compartment (transwell) of a transwell plate for detection of apoptosis and necrosis. Neutrophils cultured in the presence of LPS were used as a positive control. Co-cultured neutrophils were collected as above and cell death was quantified with flow cytometry. (**p < 0.01, vs resting neutrophils alone).