Fig. 2

AGEs upregulate β-catenin protein level and activate its transcriptional activity. A The enrichment analysis of KEGG pathways of differentially expressed genes (DEGs) induced by AGEs. B Heatmap of Representative DEGs enriched in the Wnt signaling pathway. C qPCR analysis of representative DEGs enriched in Wnt signaling pathway. n = 4. D HUVECs were treated by AGEs (100 μg/mL) for different times and β-catenin protein level was measured by western blot. n = 5. E qPCR analysis of β-catenin mRNA level in HUVECs with AGEs (100 μg/mL) treatment for different times. n = 4. F Heatmap of Wnt signaling upstream genes that affect β-catenin protein level, including Wnt receptors, downstream genes of them and destruction complex genes. G qPCR analysis of Wnt signaling upstream genes mRNA levels in HUVECs with AGEs (100 μg/mL) treatment. n = 4. H HUVECs were treated by AGEs (100 μg/mL) for different times. Proteins of nuclear fractions were isolated and then β-catenin protein was detected by western blot. Lamin B1 was used as an internal control for nuclear proteins. n = 5. NL indicates the nuclear lysate. I HUVECs were treated by AGEs (100 μg/mL) for 8 h, and then stained for β-catenin (red). The nuclei were stained with DAPI (blue). The line charts show the mean fluorescence intensity (MFI) of the distance in the images from α to ω in arbitrary units (AU). Scale bar indicates 50 μm. J 293T cells were transfected with luciferase constructs containing TCF/LEF response element (RE) and cultured for 24 h followed by AGEs treatment. Dual-Luciferase Reporter Assay was performed to detect the luciferase reporter activities the and luminescence was normalized to Renilla. n = 8. Data are shown as Mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001