Project description:To screen genes regulated by tachykinin in the 2-weeks mouse ovary, we have performed microarray expression profiling. COX-2 gene expression was upregulated in the mouse ovary treated with tachykinin receptor agonist compared to the mouse ovary treated with tachykinin receptor antagonists.
Project description:To investigate the similarity of toll-like receptor tolerance in macrophages stimulated with different toll-like receptor ligands we stimulated naïve or tolerant macrophages with ligands for TLR4, TLR2, TLR3 and TLR9. The data identifies a core set of genes that are tolerised by all ligands and genes that show TLR specific patterns.
Project description:Damaging STAB2 gene variants are associated with increased venous thromboembolic risk. STAB2 encodes stabilin-2, a clearance receptor, expressed by the liver and spleen. Given its function, it is likely that the prothrombotic state associated with stabilin-2 deficiency is due to reduced procoagulant protein clearance, but the identity of these ligands is unknown. We aimed to identify plasma stabilin-2 ligands using proximity biotinylation proteomics. Cells stably expressing stabilin-2-TurboID were incubated with human plasma and biotin to initiate TurboID labeling of plasma ligands in endocytic vesicles. Biotinylated proteins were purified and identified using mass spectrometry. Candidate plasma ligands with roles in hemostasis were fluorescently labeled and incubated with stabilin-2 expressing and control cells. Flow cytometry assessed ligand surface binding and confocal microcopy assessed colocalization with stabilin-2 and lysosomes. Furthermore, plasma levels of these ligands were measured in Stab2 deficient mice and littermate controls. Twenty-eight stabilin-2 specific ligands were identified. Interactions with von Willebrand factor (VWF), fibrinogen, pro(thrombin), heparin cofactor II (HCII), high molecular weight kininogen (HMWK), plasminogen and C4b binding protein (C4bp) were probed. HCII, HMWK, plasminogen and fibrinogen showed binding to stabilin-2 using flow cytometry (>2-fold higher than controls). Confocal microscopy demonstrated stabilin-2 dependent colocalization of all ligands with lysosomes. In Stab2 deficient mice, ligand levels were not significantly higher than wild type, suggesting in mice stabilin-2 is not their main clearance receptor. These results confirm the value of proximity labeling proteomics in identifying receptor ligands and suggest damaging STAB2 variants increase venous thromboembolic risk through altered clearance of hemostatic proteins.
Project description:Analyzed differentially expressed genes among FOP- or resFOP-iMSCs treated by several ligands: Activin-A, 100 ng/mL; BMP-7, 100 ng/mL; TGF-B3, 10 ng/mL Comparison of gene expressions among FOP- or resFOP-iMSCs treated 16h by several ligands
Project description:The macrophage mannose receptor (CD206, MR) is an endocytic lectin receptor which plays an important role in homeostasis and innate immunity. The MR contains C-type lectin domains (CTLD) whose specificity for endogenous glycans and glycoprotein ligands have not been well studied. To gain more insights into the recognition by the MR, we used the murine MR CTLD 4-7 coupled to the Fc-part of IgG (MR-Fc) to investigate its glycan and glycoprotein recognition. As lung cancer tissue and the lung cancer cell line A549 showed intense MR-Fc binding, we further investigated the MR glycoprotein ligands in those cells by immunoprecipitation and glycoproteomic analysis. All enriched glycoproteins, of which 42 were identified, contained pauci- or oligomannose N-glycans, confirming the microarray results. Our study demonstrates that the MR CTLD4-7 is highly selective for pauci- and oligomannosidic N-glycans, structures that are often elevated in tumor cells, and suggest a potential role for the MR in tumor biology.