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. 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 3: NETs decrease integrin-mediated efferocytosis.

Figure 3:

(A, B) Peritoneal macrophages (5 × 105) and CFSE-labeled apoptotic cells (1.5 × 106) in the presence of NETs (1000 ng/mL) with or without rmMFG-E8 (2 μg/mL). After 1 h of incubation, the cells were collected, washed, and stained with PE-F4/80 Ab and assessed efferocytosis in macrophages by flow cytometry. Data were obtained from 3 independent experiments and expressed as means ± SE (n=6 mice/group). The groups were compared by one-way ANOVA and SNK method. *p<0.05 vs. NETs(−) rmMFG-E8(−); #p<0.05 vs. NETs(−) rmMFG-E8(+). (C-F) Impaired surface expression of integrins in NETs-treated macrophages. Peritoneal macrophages (5 × 105) were treated with NETs at various doses. After 1 h of stimulation, the macrophages were washed, stained with (C, D) anti- αvβ3 and (E, F) αvβ5 integrin Abs and detected integrins’ expression by flow cytometry. Data were obtained from 3 independent experiments and expressed as means ± SE (n=9 samples/group). The groups were compared by one-way ANOVA and SNK method. *p<0.05 vs. PBS-treated macrophages. rmMFG-E8, recombinant mouse milk fat globule-EGF-factor VIII.