Fig. 4
Effect of rA2 on TBI-induced neutrophil infiltration and activation in the mouse brain. A Single-cell suspensions were prepared from brain tissues of mice with TBI at 24 h after surgery. Flow cytometry plots show gating strategy of brain-infiltrated immune cells including microglia (CD11b+CD45int), neutrophils (CD11b+CD45highLy6G+), monocyte/macrophages (CD11b+CD45highF4/80+), and T lymphocytes (CD45highCD3+), and the expression of IL (interleukin)-6, TNFα (tumor necrosis factor-α), and IL-10 in neutrophils. All gates were set using FMO controls. B Quantitative analysis of CNS-invading leukocytes in the brains of sham, TBI, and TBI + rA2 mice, n = 10. C Quantitative analysis of microglia in the brains of sham, TBI, and TBI + rA2 mice, n = 10. D Quantification of neutrophil expressing IL-6, TNFα, and IL-10 in the brains of sham, TBI, and TBI + rA2 mice, n = 8–10. E Representative immunostaining images and quantification of CD45- and Ly6G-positive cells in the cortex area of the ipsilateral hemisphere in sham, TBI + saline, and TBI + rA2 mice brain. Scale bar, 50 μm. Data are expressed as mean ± SEM; n = 6, *p < 0.05 compared to TBI. F Representative gel images of Western blotting and quantitative analysis for Ly-6G expression in the cortex area of the ipsilateral hemisphere in C57BL6 mice at 2 days after TBI, n = 3–4. Data are expressed as mean ± SEM; *p < 0.05 compared to sham; #p < 0.05 compared to TBI