Journal: Frontiers in Immunology
Article Title: Endothelial cells regulate mesangial cells through the Dll4/Notch3 axis to participate in glomerular injury in lupus nephritis
doi: 10.3389/fimmu.2026.1720756
Figure Lengend Snippet: EC regulate MC via the Dll4/Notch3 axis. (A) ELISA assays were performed to detect the expression of Dll4 in ECs after LPS stimulation. (B) CCK8 assay showing the effects of Dll4 on MC viability. (C) qRT-PCR assay showing Notch3 relative genes were upregulated in MC after Dll4 stimulation. (D) CCK8 assay showing the effects of Tarextumab on MC viability after Dll4 interference. (E) Western blotting analysis of the expression of the Notch3-ICD and its downstream-related proteins HEY1 and HEY2 following interference with Dll4 alone or in combination with Tarextumab. (F) Quantification of Western blotting. Data were representative of three independent experiments (n = 3 per group). (G) Representative images of wound healing assays showing the migration of MC cells treated with Dll4 or Dll4 in combination with Tarextumab. Scale bar = 200 μm. (H) Quantitative comparison of MC cell migration rates among Control, Dll4, and Dll4+Tarextumab treatment groups (n = 3). (I) Transwell analysis of the effect of Dll4 and Tarextumab on MC cell migration. Scale bar = 2 mm for overview. Scale bar = 300 μm for magnified view. (J) Quantitative analysis of the number of migrated cells. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01. (K) EdU assays were performed to detect the proliferation of MC treated with Dll4 and Tarextumab. Scale bar = 200 μm. (L) Quantitative analysis of MC cell proliferation by EdU assay. Data are presented as mean ± SD (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Article Snippet: Primary antibodies used included: Notch3 (CST, 5276S, 1:1000), Hey1 (Proteintech, 19929-1-AP, 1:1000), Hey2 (Proteintech, 105791-1-AP, 1:1000), and β-tubulin (CST, #2128S, 1:1000).
Techniques: Enzyme-linked Immunosorbent Assay, Expressing, CCK-8 Assay, Quantitative RT-PCR, Western Blot, Migration, Comparison, Control, EdU Assay