Moreover, activation of β-catenin was shown to regulate the local immunity and tolerance balance in murine intestinal mucosa.16 Despite its essential immunomodulatory www.selleckchem.com/products/cx-5461.html functions, however, little is known of the molecular mechanisms by which β-catenin may regulate DC function and/or local inflammation

responses in the liver. Here we report on the crucial regulatory function of STAT3-induced β-catenin on DC function and inflammatory responses in hepatic IRI. We demonstrate that β-catenin inhibits phosphatase and tensin homolog delete on chromosome 10 (PTEN) and promotes the PI3K/Akt pathway, which in turn down-regulates DC immune function and depresses TLR4-driven inflammation. Our data document β-catenin as a novel regulator of innate and adaptive immune responses in the mechanism of liver IRI. Ad-β-gal, recombinant

adenovirus β-galactosidase reporter gene; BMDCs, bone marrow derived-dendritic cells; DC, dendritic NVP-LDE225 clinical trial cell; GSK-3β, glycogen synthase kinase 3β; HO-1, hemeoxygenase-1; IRF3, interferon regulatory factor-3; LPS, lipopolysaccharide; PI3K, phosphoinositide 3-kinase; PTEN, phosphatase and tensin homolog delete on chromosome 10; sGPT, serum glutamic-pyruvic transaminase; siRNA, small interfering RNA; TLR4, Toll-like receptor 4; TUNEL, terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end labeling. Male C57BL/6 wildtype (WT) mice at 6-8 weeks of age were used (Jackson Laboratory, Bar Harbor, ME). Animals, housed in UCLA animal facility under specific pathogen-free conditions, received humane care according to the criteria outlined in the “Guide for the Care and Use of Laboratory Animals” (NIH publication 86-23 revised 1985). Murine BMDCs and liver DCs were generated as described.17, 18 In brief,

bone-marrow cells from femurs of WT mice were MCE cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS), 100 μg/mL of penicillin/streptomycin (Life Technologies, Grand Island, NY), in 12-well plates (1 × 106 cells/mL) with granulocyte-macrophage colony-stimulating factor (GM-CSF, 20 ng/mL, R&D Systems, Minneapolis, MN) and IL-4 (10 ng/mL, R&D Systems). Adherent immature DCs (purity ≥90% CD11c+) were recovered for in vitro experiments on day +7. To separate hepatic DCs, mouse livers perfused with phosphate-buffered saline (PBS) followed by collagenase type IV/DNase 1 (Sigma-Aldrich, St. Louis, MO). After washing, the resuspended cells were incubated with antimouse CD11c-coated immunomagnetic beads (Stemcell Technologies) for 15 minutes at 4°C and positively selected by using a magnetic column according to the manufacturer’s instruction. For DC maturation studies, CD11c-enriched cells were cultured for 24 hours with lipopolysaccharide (LPS; 0.5 μg/mL). siRNA against β-catenin was designed using the siRNA selection program.