Fig. 6: Administration of the CBL-b C373 peptide mitigated VD3-induced VC.
From: CBL-b E3 ligase-mediated neddylation and activation of PARP-1 induce vascular calcification
a Experimental procedure for administering the C373 peptide (1 mg/kg/day) via intraperitoneal injection to mice with VD3-induced VC. b Alizarin red S staining of the entire aorta in the mice. Scale bar, 10 mm. Aortic calcium (c) and serum calcium (d) levels were measured. Administration of the C373 peptide reduced VD3-induced calcium accumulation. n = 6 per group, independent experiments. e Immunoprecipitation with an anti-NEDD8 antibody was used to detect PARP-1 neddylation. PARP-1 neddylation was diminished by administration of the C373 peptide to the aortas of VD3-induced VC mice. f PARP-1 activity was detected by the PAR polymer with a boronate pull-down assay. The C373 peptide blunted VD3-induced PARP-1 activity. g Histology: alizarin red S staining (left panel scale bar, 200 μΜ) and PLA with anti-NEDD8 and anti-PARP-1 (right panel, scale bar, 20 μΜ) were performed on cross-sections. VD3 increased vascular calcification, but was inhibited by the C373 peptide. The administration of the CBL-b C373 peptide led to the dissociation of NEDD8 from PARP-1. “*” indicates a nonspecific band. The data are presented as the means ± SEMs. Statistical significance was tested via ANOVA with Tukey’s HSD test and Dunnett’s T3 test.
