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Blood, Vol. 112, Issue 8, 3175-3185, October 15, 2008
Hepatic microenvironment programs hematopoietic progenitor differentiation into regulatory dendritic cells, maintaining liver tolerance
Blood Xia et al.
112: 3175
Supplemental materials for: Xia et al
Files in this Data Supplement:
- Figure S1. Characteristics of liver stromal cells prepared form neonatal C57BL/6 mice (JPG, 96 KB)
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(A) Liver stromal cells stained with anti-Albumin (specific for hepatocyte), anti-Cytokeratin-7 (specific for biliary epithelial cell), anti-Vimentin (specific for liver fibroblast), anti-Desmin(specific for liver fibroblast), and then PE conjugated goat anti-mouse IgG, were detected by fluorescence microscopy and FACS (Left panel). For flow cytometery analysis, open histograms represent isotype control; numbers in histograms indicate the geometric mean fluorescence. To distinguish from liver sinusoidal endothelial cells which have potent phagocytosis ability for Dil-Ac-LDL(acetylated low-density lipoprotein labeled with 1,1′–dioctadecyl-3,3,3′,3′–tetramethylindo-carbocyanine perchlorate, Biomedical Technologies Inc.), liver stromal cell were treated with Dil-Ac-LDL and then detected under fluorescence microscope. Macrophages were used as a positive control. Original magnification, ×400. (right panels). (B) Phenotype of liver stromal cells.(C) Fibroblast characteristics of liver stromal cells by RT-PCR. Chemokine (D) and cytokine (E) profiles of liver stromal cells were detected by RT-PCR. Data are representative of at least three independent experiments.
- Figure S2. Functional characteristics of LRDCs natural counterpart in liver (JPG, 95.5 KB)
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(A) IL-10 and IL-12p70 secretion by the sorted CD11clowIA-blowCD11bhi cells stimulated with 100 ng/ml LPS were assayed by ELISA. (B,C) The inhibitory effect of the sorted CD11clowIA-blowCD11bhi cells on cDCs-induced OVA-specific CD4 T cell proliferation. The living CD4 T cells were counted by FACS (B) and the cytokine secretion was detected by ELISA (C). (D) Apoptosis assay in CD11clowIA-blowCD11bhi cells/cDCs/CD4 T cells coculture system by Annexin V staining. Data present as mean ± SD of triplicate samples and are representative at least from three independent experiments. *: P<0.001.
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