Figure 4. Effect of CD44 on macrophage migration.

A) Monocyte-derived macrophages were incubated in the absence or presence of CD44 mAb for 30 min to binding prior to “wounding” of the cell monolayer. Assessment of migration into the wounded area was made by microscopy at 18 h, data shown are mean number of cells present in the wound ± SEM from 6 separate experiments. In paired analysis there is a significant reduction of migration in the presence of CD44 mAb (p<0.05). B) Monocyte-derived macrophages were incubated in the absence or presence of CD44 mAb for 30 min to binding prior to “wounding” of the cell monolayer. Assessment of macrophage migration into the wounded area was made by quantification of the numbers of macrophages in the wound using time lapse microscopy images captured over 10 hours. Measurements for untreated macrophages are indicated by gray diamond symbols and CD44-treated macrophages by black squares. Data are mean number of cells observed in the wound ± SEM from 3 independent experiments. * indicates that migration was significantly reduced by CD44 mAb from 3 h onwards (p<0.05). Monocyte-derived macrophages adherent to glass coverslips were treated with media (C) or CD44 mAb (D) for 30 min prior to fixation with paraformaldehyde and staining for filamentous actin with rhodamine phalloidin. Representative micrographs show localisation of podosome-like structures in adherent macrophages (C) and marked redistribution that is observed following CD44 antibody binding (D). Scale bar = 10 µm.