Figure 2.

Shp-2 recruitment to coated beads. A: HUVEC monolayers were exposed to polystyrene beads coated with non-specific mouse IgG (ns-mIgG), DNAM-1-Fc chimera, or antibodies against PVR or PECAM for 20 minutes and fixed. Shp-2 and PECAM were visualized using immunofluorescence, as indicated. Beads for the corresponding images were visualized using bright field microscopy. Arrows, beads with Shp-2 recruitment. Shp-2 recruitment around beads was variable, depending on its position on the cell engaged and the length of time the bead had bound. Scale bar = 10 μm. B: The number of beads showing enrichment of Shp-2 fluorescence, as seen in A, was scored from five experiments in which approximately 50 beads were scored at random for each sample. Enrichment was defined as those beads with at least a 1.5-fold increase in intensity in the area immediately surrounding the bead of that compared with the surrounding cytosol. Data shown are the mean and SD from five independent experiments. *P < 0.05, determined using the Student’s t-test for unpaired observations.