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. 2010 Jun 8;123(13):2308–2318. doi: 10.1242/jcs.058321

Fig. 2.

Fig. 2.

Pro-HB-EGF is localized to sites of cell-cell contact. After 24 hours of transfection of COS-7 cells with AP–HB-EGF–GFP, the biotinylated extracellular acceptor peptide in AP–HB-EGF–GFP was labeled with streptavidin-Cy5 (red) and imaged alongside the cytoplasmic tail conjugated to EGFP (green), N-cadherin immunostaining (white) and phase contrast (A) in a confluent monolayer, (B) after 4 hours of wound healing and (C) in sparsely plated cells. Both the extracellular and intracellular domains of HB-EGF were localized to sites of cell-cell contact (arrowheads) and absent from free edges. In B, cells are migrating from left to right to close the wound. Each row represents the same field. Scale bars: 40 μm. (D) The fraction of HB-EGF colocalized with N-cadherin is relatively constant between cells that were sparsely plated (triangles), at the wound edge (circles) or in a confluent monolayer (squares), despite different values for the fraction of the cell perimeter in cell-cell contact. Data shown are average and standard deviation for n≥8.