FIG 5.

LecB inhibits epithelial wound healing. (A to C) Polarized monolayers of MDCK cells grown in 12-well plates were wounded with a pipette tip and imaged with a wide-field microscope at the indicated time points to observe wound closure. In panel A, cells were treated with LecB and/or l-fucose (43 mM) to block LecB, whereas in panel B increasing concentrations of LecB were used. The quantification of the migration speeds of the wound edges from the latter experiment (C) shows that concentrations larger than 50 μg/ml LecB completely inhibit wound healing. n = 3. (D) Polarized monolayers of MDCK cells stably expressing the plasma membrane marker ML-GFP (green) grown on chambered cover glasses were wounded and left untreated (ctrl) or treated with LecB followed by live imaging of the wound edge by confocal microscopy. Lamellipodia are indicated with arrows. (E) Polarized MDCK monolayers grown on chambered cover glasses were wounded and treated with LecB-Alexa Fluor 488 (green) for 3 h. Cells were fixed and stained for β1-integrin (red). An x-y confocal section at half height of the cells is shown. Arrows point to internalized β1-integrins colocalizing with LecB-Alexa Fluor 488; the dashed line outlines the wound edge.