Figure 5

Astrocytes reduce intracellular ROS production of neutrophils. A & C. FACS-sorted resting and LPS-primed neutrophils were stained with H2DCFDA and then were co-cultured with resting or poly I:C-primed astrocytes at a ratio of 1:1 for 6 hr. H2DCFDA intensity was then determined with a microplate reader (A) or flow cytometry analysis (C). White bar, neutrophils alone; grey 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 = 4/bar). B. Intracellular ROS production by LPS-primed neutrophils was analyzed using a transwell plate and H2DCFDA intensity was quantified by flow cytometry. Resting neutrophils were used as a negative control (*p < 0.05, primed neutrophils alone vs resting neutrophils alone; +p < 0.05, primed neutrophils plus astrocytes vs primed neutrophils alone; ++p < 0.01, transwell cultured primed neutrophils vs primed neutrophils alone. n = 4/bar). D. Representative flow cytometry graph of ROS production by neutrophils. Grey, resting neutrophils; red, LPS-primed neutrophils; blue, co-cultured primed neutrophils; brown, transwell-cultured primed neutrophils.