Fig. 3: Probing the role of ECD in stabilizing the VEGFR1 autoinhibited state.
From: Molecular basis of VEGFR1 autoinhibition at the plasma membrane
a, b Schematic representation of VEGFR1 (a) and VEGFR2 (b) constructs used in this study. c The plot of Y1175 phosphorylation level against the expression level of the constitutively activated C482R mutant of VEGFR2 in the presence or absence of VEGF165. [n = 73 (−VEGF165) and 75 (+VEGF165) cells examined over 5 independent experiments. Data are presented as mean values ± SD]. d The plot of Y1213 phosphorylation versus VEGFR1-C471R expression in the presence and absence of ligand. [n = 70 (−VEGF165) and 80 (+VEGF165) cells examined over five independent experiments. Data are presented as mean values ± SD]. e The diffusion coefficient measured from FRAP studies of indicated constructs of VEGFR1 and VEGFR2 in the presence and absence of VEGF165 are plotted. VEGFR1-GPA chimera and VEGFR1-GPA-G83I chimera represent dimer and monomer controls, respectively. Each data point in the box plot reflects the diffusion coefficient of the selected cell, and the black line indicates the mean value. n = 18 (VEGFR1-TMgPA-G83I), 24 (VEGFR1-TMgPA), 20 (VEGFR1-VEGF165), 20 (VEGFR1 + VEGF165) 24 (C471R-VEGF165), 18 (C471R + VEGF165), 25 (VEGFR2-VEGF165), 18 (VEGFR2 + VEGF165) 21 (C482R-VEGF165), and 18 (C482R + VEGF165) cells examined over 8 independent experiments. An unpaired two-tailed t-test was used to calculate significance. Boxplots represent quartiles. The data points outside the whisker range are set as outliers. The black line inside the box represents the median value. f, g The plot of the phosphorylation level of Y1175 in VEGFR2 (f) and Y1213 in VEGFR1 (g) against the indicated receptor expression level in the presence and absence of the ligand. In panel f, n = 72 (VEGFR1-VEGF165), 70 (VEGFR1 + VEGF165), and 74 (∆ECD-VEGFR1) cells were examined over four independent experiments. In (g), n = 70 (VEGFR2-VEGF165), 62 (VEGFR2 + VEGF165), and 91 (∆ECD-VEGFR2) cells were examined over four independent experiments. Data points are represented as mean values ± SD. All data were plotted using GraphPad Prism Ver 9.5.1. The boxplots were generated using Origin Pro 2020b. All the schematics and icons were designed using Inkscape Ver1.2. Source data are provided as a Source Data file for (b–g). See Supplementary Figs. 3, 4.
