Fig. 1: Characterization of the lateral diffusion of ENG, NRP1 and VEGFR2.
From: NRP1 interacts with endoglin and VEGFR2 to modulate VEGF signaling and endothelial cell sprouting
COS7 cells were transfected by vectors encoding HA-ENG, myc-NRP1 or myc-VEGFR2. At 24 h post-transfection, the cell-surface receptors on live cells were labeled at 4 °C by monovalent fluorescent Fab’ fragments (see Methods), and subjected to FRAP studies conducted at 15 °C to minimize internalization (Methods). Representative FRAP curves showing the lateral diffusion of singly-expressed HA-ENG (a), myc-NRP1 (b), or myc-VEGFR2 (c). Solid lines represent the best-fit (by non-linear regression) to the lateral diffusion equation (see Methods). The Rf and D values for each representative experiment are shown within each panel. Average Rf (d) and D values (e) derived from multiple patch/FRAP measurements. Bars depict the mean ± SEM of multiple experiments conducted each on a different cell. Some of these numbers are lower in panel e because FRAP curves yielding less than 20% recovery can be accurately analyzed only for Rf.
