Journal: Frontiers in Molecular Biosciences

Article Title: Glycosaminoglycans and fucoidan have a protective effect on experimental glomerulonephritis

doi: 10.3389/fmolb.2023.1223972

Figure Lengend Snippet: Cortical mRNA expression levels of IL-6, ICAM-1 and Nephrin in mice with LPS-induced glomerulonephritis are affected by treatment with unstimulated HS glx , LPS HS glx , fucoidan or sulodexide. (A) . MCP-1, (B) . IL-6, (C) . IL-10, (D) . ICAM-1, (E) . VCAM-1, (F) . Nephrin, and (G) . HPSE1 cortical mRNA expression of mice injected with LPS (48 h) and treated with unstimulated HS glx , LPS HS glx , fucoidan or sulodexide. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001. n ≥ 4. Unstimulated HS glx , HS extracted from unstimulated endothelial glycocalyx; LPS HS glx , HS extracted from LPS stimulated endothelial glycocalyx; MCP-1, monocyte chemoattractant protein-1; ICAM-1, intercellular adhesion molecule 1; VCAM-1, vascular cell adhesion molecule-1; HPSE1, heparanase.

Article Snippet: Primary antibodies included rat anti-mouse Gr-1 (Ly6G) (cat#14-5931-85, Invitrogen, RRID:AB_467731), rat anti-mouse CD68 (Cat#MCA 1957, RRID:AB_322219 Serotec, Oxford, United Kingdom), rat anti-mouse ICAM-1 (Cat# 14-0541-85, RRID:AB_467302, Thermo Fisher Scientific), rabbit anti-mouse VCAM-1 (Cat# NBP2-67292, Novus Biologicals), Armenian hamster anti-mouse MI-91 (anti-agrin ( )), rabbit anti-mouse PDGFR (Cat# ab32570, RRID:AB_777165, Abcam, Cambridge, United Kingdom), and rat anti-mouse CD31 (Cat# 553369, RRID:AB_394815, BD Biosciences, Vianen, Netherlands).

Techniques: Expressing, Injection

Journal: Frontiers in Molecular Biosciences

Article Title: Glycosaminoglycans and fucoidan have a protective effect on experimental glomerulonephritis

doi: 10.3389/fmolb.2023.1223972

Figure Lengend Snippet: Endothelial ICAM-1 expression is increased in LPS-induced glomerulonephritis and is not affected by treatment with unstimulated HS glx , LPS HS glx , fucoidan or sulodexide. (A) . Expression of ICAM-1 in control kidneys and kidneys of mice with LPS-induced glomerulonephritis combined with agrin staining to visualize the glomerular basement membrane (B) . Semiquantitative analysis of the glomerular expression of ICAM-1 in control mice and mice with LPS induced glomerulonephritis, untreated and treated with unstimulated HS glx , LPS HS glx , fucoidan or sulodexide. (C) . Expression of VCAM-1 and the mesangial cell marker PDGFR shows that VCAM-1 is upregulated in mesangial cells in LPS-induced glomerulonephritis. IF staining is presented on subsequent kidney sections, in which the same glomerulus was imaged because the antibodies for VCAM-1 and PDGFR could not be combined on a single kidney section. Data are expressed as mean ± SEM. **** p < 0.0001. n ≥ 4. Unstimulated HS glx , HS extracted from unstimulated endothelial glycocalyx; LPS HS glx , HS extracted from LPS stimulated endothelial glycocalyx; ICAM-1, intercellular adhesion molecule 1; VCAM-1, vascular cell adhesion molecule-1; PDGFR, platelet-derived growth factor receptor; AU, arbitrary units.

Article Snippet: Primary antibodies included rat anti-mouse Gr-1 (Ly6G) (cat#14-5931-85, Invitrogen, RRID:AB_467731), rat anti-mouse CD68 (Cat#MCA 1957, RRID:AB_322219 Serotec, Oxford, United Kingdom), rat anti-mouse ICAM-1 (Cat# 14-0541-85, RRID:AB_467302, Thermo Fisher Scientific), rabbit anti-mouse VCAM-1 (Cat# NBP2-67292, Novus Biologicals), Armenian hamster anti-mouse MI-91 (anti-agrin ( )), rabbit anti-mouse PDGFR (Cat# ab32570, RRID:AB_777165, Abcam, Cambridge, United Kingdom), and rat anti-mouse CD31 (Cat# 553369, RRID:AB_394815, BD Biosciences, Vianen, Netherlands).

Techniques: Expressing, Staining, Marker, Derivative Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Glycosaminoglycans and fucoidan have a protective effect on experimental glomerulonephritis

doi: 10.3389/fmolb.2023.1223972

Figure Lengend Snippet: LPS-induced ICAM-1 mRNA expression in cultured mouse glomerular endothelial cells is normalized by pre-treatment with fucoidan. qPCR analysis of (A) . ICAM-1, (B) . IL-6, (C) . MCP-1 mRNA expression and protein secretion levels of (D) . MCP-1 measured in culture supernatant by ELISA of mouse glomerular endothelial cells pre-treated with 10 μg/mL unstimulated HS glx , LPS HS glx , fucoidan or sulodexide for 1 h and subsequently stimulated with 100 ng/mL LPS for 24 h. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n ≥ 3. Unstimulated HS glx , HS extracted from unstimulated endothelial glycocalyx; LPS HS glx , HS extracted from LPS stimulated endothelial glycocalyx; ICAM-1, intercellular adhesion molecule 1; MCP-1, monocyte chemoattractant protein-1.

Article Snippet: Primary antibodies included rat anti-mouse Gr-1 (Ly6G) (cat#14-5931-85, Invitrogen, RRID:AB_467731), rat anti-mouse CD68 (Cat#MCA 1957, RRID:AB_322219 Serotec, Oxford, United Kingdom), rat anti-mouse ICAM-1 (Cat# 14-0541-85, RRID:AB_467302, Thermo Fisher Scientific), rabbit anti-mouse VCAM-1 (Cat# NBP2-67292, Novus Biologicals), Armenian hamster anti-mouse MI-91 (anti-agrin ( )), rabbit anti-mouse PDGFR (Cat# ab32570, RRID:AB_777165, Abcam, Cambridge, United Kingdom), and rat anti-mouse CD31 (Cat# 553369, RRID:AB_394815, BD Biosciences, Vianen, Netherlands).

Techniques: Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay