Journal: EBioMedicine

Article Title: Soluble CLEC2 Extracellular Domain Improves Glucose and Lipid Homeostasis by Regulating Liver Kupffer Cell Polarization

doi: 10.1016/j.ebiom.2015.02.013

Figure Lengend Snippet: High expression of Clec2 in the non-parenchymal fraction of liver cells. (A) RT-qPCR of different human tissues shows CLEC2 enrichment in the testis, liver and PBL. (B) RT-qPCR of different mouse tissues. (C) Clec2 is highly expressed in non-parenchymal cells of liver compared to hepatocytes. Results are expressed as the mean ± SEM of triplicates and are representative of two independent experiments.

Article Snippet: Mouse total RNA from hepatocytes and liver non-parenchymal cells was obtained from Zen-Bio.

Techniques: Expressing, Quantitative RT-PCR

Journal: EBioMedicine

Article Title: Soluble CLEC2 Extracellular Domain Improves Glucose and Lipid Homeostasis by Regulating Liver Kupffer Cell Polarization

doi: 10.1016/j.ebiom.2015.02.013

Figure Lengend Snippet: Recombinant Fc-CLEC2(ECD) protein improved glucose homeostasis and hepatic steatosis. (A) Body weight comparison between DIO mice injected with recombinant Fc-CLEC2(ECD) protein and PBS control. (B) Recombinant Fc-CLEC2(ECD) protein reduces serum insulin concentration in DIO mice. (C) DIO mice injected with recombinant Fc-CLEC2(ECD) protein show improvement in glucose tolerance 7 days after first injection. (D) Recombinant Fc-CLEC2(ECD) protein reduces serum triglyceride concentration in DIO mice. (E) Liver triglyceride content is reduced in recombinant Fc-CLEC2(ECD) protein treated group. (n = 12) (F–I) Recombinant Fc-CLEC2(ECD) protein stimulates markers of M2 macrophages and phagocytosis in the liver and improves serum cytokine profile. (F) Up-regulation of liver macrophage and phagocytosis markers measured by RT-qPCR. (G) Liver macrophage F4/80 staining showing a wider distribution of plump clustered Kupffer cells in treated animals, with some clusters around degenerating hepatocytes and impinging on central veins, as indicated by arrows. (H) Quantification of liver macrophage F4/80 staining indicates a trended increase of F4/80 + staining in the Fc-CLEC2(ECD) treatment group. PBS control (n = 12), Fc-CLEC2(ECD) 10 mg/kg group (n = 12), Fc-CLEC2(ECD) 30 mg/kg group (n = 10). (I) Serum cytokine levels after recombinant Fc-CLEC2(ECD) protein treatment show increases in IL-13 and decreases in G-CSF, KC, and MIP-1a. *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. Results are expressed as the mean ± SEM and are representative of two independent experiments.

Article Snippet: Mouse total RNA from hepatocytes and liver non-parenchymal cells was obtained from Zen-Bio.

Techniques: Recombinant, Comparison, Injection, Control, Concentration Assay, Quantitative RT-PCR, Staining

Journal: EBioMedicine

Article Title: Soluble CLEC2 Extracellular Domain Improves Glucose and Lipid Homeostasis by Regulating Liver Kupffer Cell Polarization

doi: 10.1016/j.ebiom.2015.02.013

Figure Lengend Snippet: Effects of CLEC2 in vitro. (A–C) Activation of CLEC2 suppresses markers for M2 lineage of macrophage in vitro. (A) Anti-CLEC2 agonistic antibody induces strong aggregation of platelets. (B) Cultured RAW264.7 cells, treated with the CLEC2 agonistic antibody for 24 h, show significantly suppressed M2 marker, Arg1 , but not M1 marker, Csf2 . Antibody at 10 nM, LPS at 1 μg/ml, and IL4 at 100 ng/ml. amAb represents agonistic CLEC2 antibody. (C) Isolated peritoneal macrophages from 6 week old ob / ob animals show high expression of Clec2 but not other Clec molecules such as Clec5a , Clec7a , and Clec10a (***p < 0.001). (D) Fc-CLEC2(ECD) simulates liver oxidative phosphorylation and fatty acid oxidation in co-cultured Kupffer cells and primary hepatocytes. Co-cultured Kupffer cells and primary hepatocytes were treated with Fc-CLEC2(ECD) at a concentration of 200 ng/ml for 5 h. Fc-CLEC2(ECD) treatment induces expression of genes encoding components of oxidative phosphorylation ( Ndufs , Sdhb ) and fatty acid beta-oxidation ( Acox1 , Cpt1a ), but not fatty acid transporters ( Slc27a1 ). *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA. Results are expressed as the mean ± SEM and are representative of four independent experiments.

Article Snippet: Mouse total RNA from hepatocytes and liver non-parenchymal cells was obtained from Zen-Bio.

Techniques: In Vitro, Activation Assay, Cell Culture, Marker, Isolation, Expressing, Phospho-proteomics, Concentration Assay