Fig. 4

SETD7 regulates HG-induced endothelial injury in vitro. RAECs were cultured in different glucose concentration conditions (Control: 5.5 mM glucose + 16.5 mM mannitol, HG: 22 mM glucose) with or without siScr or siSetd7 treatment. A The protein expression of SETD7, eNOS, VE-cadherin, and VEGF-α in siScr and siSetd7 RAECs with or without HG stimulation. B The mRNA level of Setd7, Vcam1, and Enos in siScr and siSetd7 RAECs with or without HG stimulation. C Representative images of immunofluorescence staining of SETD7 and COX-2 in siScr and siSetd7 RAECs with or without HG stimulation. Scale bars, 50 μm. D Heatmap showing differentially expressed genes (DEGs) in siScr and siSetd7 RAECs with or without HG stimulation. E Venn diagram showing the identified proteins in Control with siScr vs. HG with siScr (blue, 378), HG with Setd7 vs. HG with siScr (purple, 300). A total of 78 genes were identified in three groups. F GO-based enrichment analysis of 78 DEGs in terms of the biological processes. G Representative images of wound healing assay photographed at 0, 12, and 24 h of culture (100 ×). H Representative images of the Transwell assay photographed after 36 h of culture (200 ×). RAECs were transfected with vector (Vector) or Setd7 overexpression plasmid (Setd7^oe). I The protein expression of Myc-SETD7, eNOS, VE-cadherin, and VEGF-α in RAECs transfected with Vector or Setd7^oe. J Representative images of wound healing assay photographed at 0, 12, and 24 h of culture (100 ×). K Representative images of the Transwell assay photographed after 36 h of culture (200 ×). Statistical significances were calculated using (A, B, G, I and J) two-way ANOVA, Tukey’s multiple comparisons tests, (H) one-way ANOVA, (K) two tail student’s t test. Data are expressed as the mean ± SEM, statistical analysis revealed a significant difference with ^*p < 0.05, **p < 0.01, ***p < 0.001, ###p < 0.001, ns = not significant