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. Author manuscript; available in PMC: 2023 Jul 12.
Published in final edited form as: Sci Transl Med. 2021 Jul 28;13(604):eabb1069. doi: 10.1126/scitranslmed.abb1069

Fig. 2. CLON-G–treated mouse neutrophils display a similar or even longer life span than fresh neutrophils after drug removal.

Fig. 2.

(A) Mouse neutrophils were treated with CLON-G for 3 days in vitro followed by removing CLON-G by changing the cell culture medium to normal medium (CLON-G–treated, 3 days). At the same time, freshly isolated neutrophils were cultured in the normal medium without CLON-G (Fresh mouse neutrophils). (B) Total numbers of intact neutrophils were counted at the indicated time points. All data are presented as means ± SD of three experiments. *P < 0.01. (C) Cells were stained with FITC–A-V and PI and then analyzed by flow cytometry. The percentage of healthy neutrophils was assessed accordingly. All data are presented as means ± SD of three experiments. (D) Experimental scheme for assessing the relative in vivo death rate of transplanted fresh, G-CSF alone–treated, and CLON-G–treated neutrophils in a mouse peritonitis model. TG, thioglycolate; i.p., intraperitoneal. (E) Cells in peritoneal lavage fluid were stained with PE-CD45.1 and analyzed by flow cytometry. (F) Flow cytometry analysis of donor neutrophils in peritoneal lavage in mice transplanted with indicated neutrophil populations at each time point. Shown are representative flow cytometry plots from one of three independent experiments. (G) The ratio of indicated transplanted neutrophil populations in peritoneal lavage fluid. All data are presented as means ± SD of three experiments. *P < 0.01 compared to 0 hours.