Fig. 5.

3-D migration of peritoneal macrophages along MCP-1 and RANTES gradients. A Schematic drawing of experimental setup within a Corning transwell insert, with yellow arrows indicating the direction of macrophage migration. WT macrophages were labeled red (PKH26) and α7nAChR^−/− macrophages were labeled green (PKH67) before being added to the membrane. Migration was initiated using RANTES (12.8nM) or MCP-1 (30nM) in medium added to the top of the fibrin gel. B 3-D view of labeled macrophages migrating inside the fibrin gel after 48 h. C IMARIS 8.0 reconstruction of WT (red) and α7nAChR^−/−(green) macrophages before the initiation of migration and after 48-h incubation. Left shows the top view of individual and combined channels. Center, side view showing starting point at 0h. Right, side view showing macrophages migrating along MCP-1 or RANTES gradients. D The number of macrophages migrating greater than 80μm was analyzed as a percentage of WT (MCP-1, n = 4; RANTES, n = 3). Statistical analysis was carried out using a student's t test. *P < 0.05