Figure 5
AA-induced modulation of macrophage phenotype in vivo and effects on macrophages in vitro. (A) Flow cytometry analysis of leukocyte/lymphocyte (CD45+) classically activated macrophages (CD45+CD11b+F4/80+CD206−Cx3CR1+), alternatively activated macrophages (CD45+CD11b+F4/80+CD206+Cx3CR1+), CD3+ cells (CD45+CD3+), CD4+ T-cells (CD45+CD3+CD4+CD8−), CD8 + T-cells (CD45+CD3+CD4+CD8+) and neutrophils (CD45+CD11b+Ly6G+) after acute and chronic AAI-induced kidney injury. (B) Cell viability of macrophages was assessed by PI staining and LDH assay up to 96 hours in medium supplemented with 2% FCS. Data represents mean values 72 hours after stimulation with indicated AAI concentrations ± SEM of three independent experiments; *p < 0.05 **p < 0.01, ***p < 0.001. (C) Macrophages were stimulated with 50 µM AAI for 24 hours and stained with AnnexinV (AnV) and PI. Early apoptotic (AnV+ PI−) and late apoptotic/primary necrotic (AnV+ PI+) macrophages were identified by flow cytometry. Data are expressed as means ± SEM; ****p < 0.0001, AAI versus medium control group.
