In vivo evidence that the LMIR5–TIM1 interaction induced accumulation of neutrophils. (A and B) Relative gene expression levels of TIM1 (left) or LMIR5 (right) in the IRI or contralateral (Co) kidneys from WT mice at different time intervals after surgery (A) or in the CD45⁻ or CD45⁺ cells sorted from the IRI kidneys of WT mice at 24 h after surgery (B). Data are means ± SD (n = 6 mice in each group). Data are representative of three independent experiments. (C) Surface expression levels of LMIR5 (bottom, continuous line histograms) as well as CD11b and Gr-1 (top) were examined in BM neutrophils from WT or LMIR5^−/− (KO) mice. Control staining with goat IgG is shown (shaded histograms). Data are representative of five independent experiments. (D) Percentages of CD11b⁺Gr-1⁺ neutrophils in either contralateral or IRI kidney cells from WT or KO mice at 24 h after surgery (left). The ratio of neutrophil counts in IRI kidneys to those in contralateral kidneys was determined. Data are means ± SD (n = 6 mice in each group; right). Two independent experiments were performed. (E) Either 100 µg TIM1-Fc or control Fc, or 1 mg LPS was injected into the air pouches of WT or LMIR5^−/− mice. At 4 h after injection, neutrophils recruited into the pouches were counted. Each symbol represents an individual mouse. The number of mice in each group is shown. Two independent experiments were performed. (F) IL-6 released into the dorsal air pouches (ELISA). The number of mice in each group is shown. Two independent experiments were performed. Data in D–F are means ± SD. Statistically significant differences are shown. *, P < 0.05.