Fig. 3

α7nAChR deficiency impedes the migration of macrophages to organs during LPS-induced endotoxemia. A Schematic representation of the experimental design. Monocytes were isolated from bone marrow of male WT and α7nAChR^−/− mice via MACS. Cells were labeled with red (WT) or green (α7nAChR^−/−) fluorescent dyes, mixed in equal proportion and injected in tail vein of male WT recipient mice. After 48 h, the lung, liver and spleen were isolated, digested and analyzed using flow cytometry. B Representative dot plots of monocyte purity analysis. Isolated monocytes were labeled with anti-CD11b (APC) and anti-Ly6-G (FITC). Monocyte population is visible in Q2. C Representative results of flow cytometry analysis are shown. The leukocyte distribution in lungs before the adoptive transfer (Upper panel) and at 48 h after adoptive transfer and LPS administration (Lower panel) are presented. Data were analyzed using FACSDiva software. Migrated WT macrophages (red) were detected in Quadrant 4; α7nAChR^−/− macrophages (green) were detected in Quadrant 1. D Imaging flow cytometry of labeled macrophages. (BF = bright field, SSC = side scattering). E Bar graphs representing the amount of WT and α7nAChR-deficient macrophages detected in lungs, liver, and spleen by flow cytometry, (n = 6). Statistical analysis was performed using student’s t test. *P < 0.05, **P < 0.01