Fig. 5. Old Rragc^S74N/+ organs promote neutrophil infiltration.

a, Quantification of IHC staining for myeloperoxidase in kidneys (left) and livers (right) collected from 18-mo-old Rragc^+/+ (n = 8) and Rragc^S74N/+ (n = 7) male mice. b, Reactive oxygen species production by neutrophils isolated from Rragc^+/+ (n = 4) and Rragc^S74N/+ (n = 4) BM as assessed by flow cytometry with DHR 123 after a 20-min treatment with PMA at the indicated concentrations. c, Kinetics of phagocytosis of GFP⁺ E. coli expressed as a percentage of GFP⁺ cells by neutrophils isolated from Rragc^+/+ (n = 3) and Rragc^S74N/+ (n = 3) BM. Data are presented as mean ± s.d. d, Experimental setup of the neutrophil adoptive transplant experiment. BM-derived neutrophils were isolated from 2–3-mo-old Rragc^+/+ mice (CD45.1.1) and transplanted to 3-mo-old Rragc^+/+ (CD45.2.2) and to 3-mo-old Rragc^S74N/+ (CD45.2.2) male hosts. e, Quantification of wt CD45.1.1 neutrophils in the indicated tissues from CD45.2.2 Rragc^+/+ (n = 5) and Rragc^S74N/+ (n = 6) male hosts 16 h after the adoptive transplant. f, Quantification of CD45.1.1 wt neutrophils by IF in kidneys from CD45.2.2 Rragc^+/+ (n = 4) and Rragc^S74N/+ (n = 6) host males. g, Experimental setup of the neutrophil adoptive transplant experiment. BM-derived neutrophils were isolated from 3-mo-old Rragc^+/+ mice (CD45.1.1) and transplanted to 12–14-mo-old Rragc^+/+ (CD45.2.2) and Rragc^S74N/+ (CD45.2.2) male hosts. h, Quantification of transferred wt CD45.1.1 neutrophils in the indicated tissues from CD45.2.2 Rragc^+/+ (n = 3) and Rragc^S74N/+ (n = 3) male hosts 16 h after adoptive transplant. i, Quantification of transferred wt CD45.1.1 neutrophils by IF in kidneys from CD45.2.2 Rragc^+/+ (n = 3) and Rragc^S74N/+ (n = 3) host males. Statistical significance was assessed by two-tailed Student’s t-test (a–c,f,i) and two-way ANOVA (e,h). Data in b,c,e,f,h,i, are presented as mean ± s.d.

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