Fig. 4.

Irf8 deletion triggers neutrophil recruitment and NET formation during AKI. (A) Study design. Irf8^+/+ and Irf8^−/− mice were subjected to IR surgery and sacrificed 1 day after AKI. The kidneys and spleens of mice were used for analysis. (B) Flow cytometric analysis of kidney and spleen neutrophils (CD45⁺CD49⁻CD3e⁻CD19^-Ly6g⁺CD11b⁺, gated as indicated in Supplemental Fig. 2A). (B′-B″) The percentages and numbers of kidney/spleen neutrophils from healthy controls and mice after AKI (n = 4–5 mice per group). (C–C′) Distribution and cell numbers of Ly6g⁺ neutrophils in kidneys and spleens (n = 4–5 mice per group) from healthy controls and mice after AKI. (D) Distribution of myeloperoxidase (MPO) in kidneys (n = 4–5 mice per group). Scale bar, 200 μm. (D′) Numbers of MPO⁺ cells in kidneys. (E-G) Serum was collected from healthy and AKI mice, and the release of MPO, citrullinated histone 3 (Cit-H3), and neutrophil elastase (NE) was determined using ELISA kits (n = 5 mice per group). Each dot represents one mouse. Red star and blue arrow indicate positive cells. The data are shown as the mean ± SD. ***p < 0.001, ****p < 0.0001. Two-way ANOVA. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)