Skip to main content
. Author manuscript; available in PMC: 2022 Feb 15.
Published in final edited form as: J Immunol. 2020 Dec 30;206(4):797–806. doi: 10.4049/jimmunol.2000091

Figure 5: NE inhibitor reverses NET-dependent impairment of efferocytosis.

Figure 5:

(A, B) Murine peritoneal macrophages (5 × 105) and CFSE-labeled apoptotic cells (1.5 × 106) in presence of NETs (1000 ng/mL) and various doses of NE-I. After 1 h of cell culture, the cells were washed and stained with PE-F4/80 Ab and assessed efferocytosis using flow cytometry. Data were obtained from 3 independent experiments and expressed as means ± SE (n=8 samples/group). The groups were compared by one-way ANOVA and SNK method. *p<0.05 vs. NETs(−) NE-I(−); #p<0.05 vs. NETs(+) NE-I(–). (C) Murine peritoneal macrophages (5 × 105) were plated in 24-well plate. CFSE-stained apoptotic cells (1.5 × 106) were added to the macrophages. The cells were treated with NETs (1000 ng/mL) and various doses of NE-I. After 1 h of incubation, media was carefully removed, washed the cells with PBS, followed by fixing and stained the cells with PE-F4/80 Ab. Efferocytosis was detected by using fluorescent microscope at ×400 original magnification. Representative images in each group were chosen from 10 randomly captured images. Scale bar: 40 μm.