BMDM were infected (La) or not (control) with L. amazonensis axenic amastigotes for 1h. BMDM were then washed to remove extracellular parasites, cultured at 34°C for another 6, 24 or 48 h and assayed for their ability to migrate. For the Boyden chamber chemotaxis migration assays, macrophages were cultured at 34°C for another 5 h. (A) Effect of L. amazonensis infection in the migration velocity of macrophages, measured by phase-contrast time-lapse imaging. The data are representative of nine independent experiments. (B) Tracks representing the migration of individual macrophages, non-infected or infected for 48 h, derived from stacks of confocal images acquired every min for 1 h. The results of nine independent experiments are shown for each condition. (C) Effect of L. amazonensis infection in macrophage migration through a Boyden chamber membrane containing MCP-1 in the lower chamber. The data are representative of three independent experiments. * p < 0.05, ** p < 0.01, ** p < 0.001 (Student’s t test) compared with non-infected control (C).