Fig. 7

VEGFR1 isoform 5 accounts for β-catenin Y142 phosphorylation. A Venn diagram showing overlapping genes between the kinases of β-catenin Y142 predicted in silico from the GPS database [34] and AGE-upregulated kinases from DEGs of transcriptomic. B Schematic diagram of the 7 isoforms of VEGFR1. Detailed information was obtained from the Uniprot database. C HUVECs were stimulated by AGEs (100 μg/mL) for 1 h, and the full-length VEGFR1 levels were measured by western blot. n = 5. D HUVECs were treated with AGEs (100 μg/mL) for 1 h and then Co-IP assay was performed with β-catenin antibody to analyze the interaction between β-catenin and the full-length VEGFR1. Experiments were repeated three times with similar results and one representative result is shown. E HUVECs were stimulated by AGEs (100 μg/mL) for 1 h, and the VEGFR1 isoform5, isoform6 and isoform7 protein levels were measured by western blot. n = 8. F HUVECs were stimulated by AGEs (100 μg/mL) for 1 h and then Co-IP assay was performed with β-catenin antibody to analyze the interaction between β-catenin and isoform5/6/7. Experiments were repeated six times with similar results and one representative result is shown. G The HA-tagged VEGFR1 isoform5, isoform6 or isoform7 plasmid was transfected into 293T cells followed by AGEs treatment for 1 h, and then Co-IP assay was performed with HA antibody to analyze the interaction between endogenic β-catenin and ectogenic VEGFR1 isoform5/6/7. Experiments were repeated three times with similar results and one representative result is shown. H The Flag-tagged β-catenin-overexpressing plasmid was co-transfected with HA-tagged VEGFR1 isoform5, isoform6 or isoform7 plasmid into 293T cells followed by AGEs treatment for 1 h, and then Co-IP assay was performed with HA antibody to analyze the interaction between ectogenic β-catenin and these three isoforms of VEGFR1. Experiments were repeated three times with similar results and one representative result is shown. I HUVECs were transfected with siRNA targeting VEGFR1 isoform5 followed by AGEs (100 μg/mL) for 1 h and the phosphorylation level of β-catenin Y142 was quantified by western blot. n = 5. J 293T cells were transfected with VEGFR1 isoform5-overexpression plasmid followed by AGEs (100 μg/mL) for 1 h and the phosphorylation level of β-catenin Y142 was quantified by western blot. n = 5. Data are shown as Mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001