Journal: Frontiers in Physiology
Article Title: A new SO 2 probe ZSO targeting VDBP inhibits high glucose induced endothelial cell senescence and calcification
doi: 10.3389/fphys.2025.1719853
Figure Lengend Snippet: ZSO inhibits high glucose-induced vascular endothelial cell senescence and calcification through downregulating VDBP protein levels. (A) Endothelial cells were treated with high glucose (30 mM) and various concentrations of ZSO (1, 5, 10 μM) for 24 h. RT-qPCR was then performed to quantify VDBP mRNA level, and the results were statistically analyzed. (B) Endothelial cells were treated with DOM and various concentrations of ZSO (1, 5, 10 μM) for 4 days. RT-qPCR was then performed to quantify VDBP mRNA level, and the results were statistically analyzed. (C) Endothelial cells were treated with high glucose (30 mM) and ZSO (5 μM) for 24 h, followed by Western blot analysis of VDBP protein level. (D) Statistical analysis of VDBP protein level under senescence conditions. (E) Endothelial cells were treated with 5 μM ZSO under calcification-inducing conditions for 4 days, followed by Western blot analysis of VDBP protein level. (F) Quantitative analysis of VDBP protein level under calcification conditions. (G) Immunohistochemical staining of VDBP in mouse thoracic aorta sections, with brown coloration indicating positive immunoreactivity. (H) Quantitative analysis of VDBP-positive staining intensity in vascular tissues, with corresponding statistical graphs. (I) Measurement of VDBP protein level in mouse serum using an ELISA kit, followed by statistical analysis. (J) Endothelial cells were treated with high glucose (30 mM) and 5 μM ZSO for 24 h, and the ubiquitination level of VDBP was assessed. (K) Quantification of VDBP ubiquitination level. ns p > 0.05, *p < 0.05, **p < 0.01, n = 3.
Article Snippet: Beads were washed, and Western blot was used for detection, first incubated with ubiquitination antibody (Abcam, ab137031), developed, and then incubated with VDBP antibody (Santa Cruz, sc-365441) and developed.
Techniques: Quantitative RT-PCR, Western Blot, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay, Ubiquitin Proteomics
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