These findings indicate that diet composition can

significantly influence the composition of Emu Oil27 and hence possibly impact on oil efficacy.30 In a CD-1 mouse model of croton oil-induced auricular inflammation, topical CP-690550 price application of Emu Oil significantly decreased auricular thickness and weight.31 Furthermore, Emu Oil reduced levels of the pro-inflammatory cytokines TNF-α and IL-1α,31 cytokines also reported to be directly involved in the development of IBD.32–34 Interestingly, the anti-inflammatory effects of Emu Oil in croton oil-induced auricular inflammation were more pronounced than application of fish, flaxseed and olive oils, or liquefied chicken fat;31 oils known to contain significantly higher levels of FAs. Snowden and Whitehouse23 assessed the anti-inflammatory activity of five different preparations of Emu Oil, varying in Emu farm location, source of Emu adipose tissue (subcutaneous or retroperitoneal), rendering protocols and storage. Five Emu Oil preparations (Emu Oil [EO] one; commercially available preparation in Western Australia [WA] with added anti-oxidant, EO two; commercially rendered in WA with no additives, EO three; prepared using intra-abdominal fat from WA birds, EO four; prepared using subcutaneous fat from Queensland birds, EO five; commercially rendered

from Queensland birds) were topically applied to rat paws, following experimentally-induced polyarthritis. Paw diameter, indicative of arthritic inflammation, was significantly reduced following application of four of the Emu Oil preparations (EO two-five). Furthermore, Emu Oil preparations two and three check details Myosin reduced inflammation to an extent comparable with oral ibuprofen (40 mg/kg), a readily available NSAID.23 Emu Oil has further been demonstrated to reduce plasma cholesterol concentrations in hypercholesterolemic hamsters compared with hamsters ingesting

a saturated fatty acid-enriched diet35 and Emu Oil administration reduced plasma low-density lipoprotein and aortic cholesterol ester concentrations.35 Whitehouse et al.22 indicated that transdermal application of Emu Oil in 15% (v/v) cineol significantly reduced paw swelling in addition to promoting weight gain in a rat model of arthritis. Bennett et al.29 demonstrated that Emu Oil has both antioxidant properties in vitro (radical scavenging activities) and a protective role against oxidative damage (assessed by measuring the ability to inhibit lipid peroxidation of erythrocytes) in a biological membrane model system. Furthermore, Emu Oil afforded greater protection against oxidative damage than the Ostrich and Rhea Oils.29 Topical application of Emu Oil has been demonstrated to promote wound healing and recovery. In a study by Politis and Dmytrowich,36 Emu Oil lotion (a mixture of Emu fat, oil, vitamin E and botanical oil) was applied to full-thickness skin defects 24 h after surgery in rodents.

2013). Sitka spruce (Picea sitchensis) infected by H. annosum-resistant clones contained considerably less fungal DNA than the susceptible ones, and host defence responses were found to be weaker in wood than in bark (Bodles et al. 2006). In general, low levels of pathogen DNA in resistant plants would characterize a mechanism that

results in the inhibition of pathogen multiplication, whereas the presence of relatively high amounts of pathogen DNA in asymptomatic plants should indicate a mechanism based on tolerance rather than on true resistance (Vandemark and Barker 2003). The fungal biomass of Alternaria dauci was equivalent in two carrot cultivars between 1 and 15 days Y-27632 cost after inoculation, while it was fourfold higher in the more susceptible cultivar 21 and 25 days after inoculation (Boedo et al. 2008). Authors speculated that the pathogen can colonize both cultivars in a similar manner during the first steps of the interaction, but fungal development is subsequently restricted in the partially resistant cultivar due to putative plant defence reactions. Instead, F. oxysporum f.sp. cubense levels in severely symptomatic banana pseudostems and leaves proved to be much higher than in Ibrutinib in vivo mild symptomatic ones (Lin et al. 2013). Furthermore, unlike the visual examination of

symptoms, the accurate quantification of the pathogen DNA could enable the detection of even minor differences in the host resistance. Blast resistance levels of rice cultivars were more accurately evaluated with qPCR, because by the time lesions on leaves had just become visible, the growth of Magnaporthe grisea was 80 times higher in susceptible than in resistant cultivar (Qi and Yang 2002). Similarly, qPCR fine-tuned

bioassays for the quantification of Cercospora leaf spot disease in sugar beet breeding (De Coninck Glutamate dehydrogenase et al. 2012). In addition, qPCR can contribute to the protection of plant species by favouring the evaluation of control measures or the selection of new anti-oomycete and antifungal compounds (Llorente et al. 2010). A specific qPCR method was developed to detect Botrytis cinerea in vineyards and utilized to compare the efficacy of different control strategies including various fungicide treatments (Diguta et al. 2010). The method could also serve as a decision-making tool in vineyards by fostering the assessment of the contamination risk and optimizing the number of sprays and the concentration of fungicides to be used. Furthermore, qPCR can be utilized to specifically monitor and quantify the frequencies of fungicide-resistant genotypes in a specific pathogen population. For example, the β-tubulin allele E198A conferring resistance to benzimidazole was quantified in Monilinia fructicola (Luo et al.

(Hepatology 2014;60:1571-1580.) Liver transplantation while patients are viremic for HCV always leads to rapid infection of the new liver. Recurrence of CHC occurs, which can affect the outcome of the transplant. HCV docks on specific surface proteins to enter into hepatocytes. Drugs impairing this binding may prevent infection. Vercauteren et al. used mice transplanted with human hepatocytes to show that the administration of monoclonal antibodies against the

scavenger receptor class B type I, which acts as an HCV receptor, successfully inhibits infection. Interestingly, this effect persisted even when the antibodies are administered after exposure to the virus and was also effective with HCV variants that are relatively resistant in Lenvatinib research buy vitro to this strategy. This is an intriguing approach that adds a new host therapeutic target and, as discussed by the investigators, it can be particularly helpful selleck chemical in the transplantation setting. (Hepatology 2014;60:1508-1518.) The discovery of the receptor for hepatitis B virus (HBV) on hepatocytes has been long awaited; its identity came as a surprise given that it is the bile acid transporter, Na+-taurocholate cotransporting polypeptide (NTCP). In a fascinating article, Oehler et al. used HBV-infected humanized mice to show that the binding of HBV to NTCP stimulates the expression of enzymes responsible for bile acid synthesis. The pre-S1-derived peptide, Myrcludex-B, which binds to both human HA-1077 and

murine NTCP, also induces the expression of enzymes responsible for bile acid synthesis. The investigators confirmed the increased messenger RNA abundance of these enzymes in liver biopsy samples from patients infected with HBV. This was accompanied by a decreased nuclear localization of the bile-acid–regulated transcription factor, farnesoid X receptor. The investigators hypothesize that these changes

occur in response to a reduction of bile acid import into hepatocytes resulting from the binding of HBV to NTCP. (Hepatology 2014;60:1483-1493.) Radiological contrast agents can provide not only morphological information, but also functional information. Magnetic resonance imaging (MRI) can be performed with contrast agents (e.g., gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid) that define arterial and venous perfusion and which are later taken up into hepatocytes by a transporter. Focal lesions composed of cells devoid of this transporter do not accumulate this contrast agent. Yamashita et al. report that the majority of HCCs do not accumulate this contrast agent, and that only well-differentiated HCCs (i.e., 15%) demonstrate uptake. These lesions are not associated with elevated alpha-fetoprotein (AFP). The investigators were able to show that the transcription factor, hepatocyte nuclear factor 4, is responsible for this phenotype. Based on an external validation cohort, they confirmed that uptake of MRI contrast agent in combination with low AFP is associated with good survival.

This fractional concentration of oxygen was maintained at this level for 60 seconds before an electric fan located at one side of the cage allowed a gradual return over 60 seconds to 20%-21% of oxygen for 60 seconds (Fig. 1). Regular checks of chamber oxygen concentrations during the experiment were made using an oxygen sensor (VMX300, Viamed, UK) PS-341 cost and were registered through an amplifier and recorded on a computer data acquisition system (ADInstruments, Mountain View, LA). For sham exposure (handled controls [HC]), rats were kept in an identical plastic cage placed side by side. The inflow gas was always room air, but the

solenoid switches, fan, and inlets reproduced the noise and airflow disturbances of the CIH protocol. Rats in both groups were housed

six per cage in accordance with space recommendations in the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Pub. No. 85-23, Revised 1985). The temperature and the relative humidity of the chambers were maintained at 21%-24°C and between 30% and 70%, respectively. At the end of the 12-hour treatment period, animals were transferred to their standard cages in a separate area and housed under MK-1775 concentration room air conditions for the remainder of the day in an animal care facility at the University of La Laguna. All protocols were approved by the Animal Care and Use Committee. Hematocrit was evaluated in control (n = 46) and CCl4-induced advanced cirrhotic rats (n = 14) to confirm the effect of CIH. The blood samples (1-2 mL) were withdrawn from the tail, placed in microcapillary tubes, and spun in a microcentrifuge (MPW-250e, Med. Instruments, Warsaw, Poland) for hematocrit measurement. At the end of the intermittent hypoxia exposure protocol, rats were anesthetized as mentioned previously. First, a catheter was inserted

in the carotid artery to monitor blood pressure (mean arterial pressure [MAP]; mm Hg) and heart rate (beats per minute, bpm), and into the portal vein through an ileocolic vein to measure portal pressure (mm Hg). The catheter was connected to a Power Lab (4SP) linked to a computer using Chart version 5.5.6 for Windows software (ADInstruments) and, after a period of 10 minutes Quisqualic acid of stabilization, recordings were performed with pressure transducers. In a supplementary group of cirrhotic rats, after baseline measurements following the stabilization period, HC and CIH rats underwent volume expansion with incremental doses (1-8 mL/kg, every 2 minutes) of hydroxyethyl starch 6% (Voluven 6% 130/0.4, Fresenius Kabi, Barcelona, Spain)16 via a femoral vein catheter. A flow-controlled perfusion system was used in this study. The system has been described elsewhere17 (see Supporting Information for details).

6C). Strikingly, EZH2-regulated miRNAs can potentially modulate cell motility-associated pathways and key signaling pathways. It was interesting to note that the first- and second-rated pathways were focal adhesion (Pathway ID hsa04510) and adherens junction (Pathway ID hsa04520), two crucial pathways in cancer cell invasion and metastasis. Consistent findings were obtained by PicTar, another miRNA target prediction algorithm (Supporting Fig. 5, Supporting Table 7). We also noticed that the RhoGTPase-associated cytoskeleton reorganization

axis was recurrently engaged selleck antibody in six of the top-rated KEGG pathways, including those for focal adhesion (Pathway ID hsa04510), adherens junction (Pathway ID hsa04520), transforming growth factor

beta (TGF-β) signaling (Pathway ID hsa04350), noncanonical Wnt singling (Pathway ID hsa04310), axon guidance (Pathway ID hsa04360), and the actin cytoskeleton regulation (Pathway ID hsa04810) (Supporting Fig. 6). We previously reported that the RhoGTPase signaling pathway is frequently altered in human HCCs and is tightly associated with HCC metastasis.21, 35, 36 Our present findings further suggest that the EZH2-tumor suppressor miRNA axis may act upstream of the pathway to mediate its perturbation. Consistent with this notion, we found that knockdown of EZH2 resulted in down-regulation of RhoA and ROCK2 protein and inhibited stress fiber formation in HCC cells (Supporting Fig. 7). Taken together, the in silico analysis reinforces the tumor suppressive functions of EZH2-regulated miRNAs, and suggests their combinational effects in modulating key cell movement selleck kinase inhibitor and metastasis-related pathways in driving STK38 HCC metastasis. Epigenetic regulation machinery involves multiple proteins with distinct functions. In our study, we first revealed that deregulation of epigenetic modifiers is common in HCCs. These epigenetic modifiers, including DNA methyltransferases, histone deacetylases, SET domain-containing histone methyltransferases, and

PcG proteins are direct mediators of epigenetic mechanisms. Their concordant deregulation reflects HCC epigenome is likely to be affected in multiple aspects. In line with our observation, not only are some of these proteins reported to be up-regulated in HCC,5, 37 but genome-wide DNA hypomethylation and promoter DNA hypermethylation of tumor-suppressors,38 as well as changes in global histone modification such as an increase of H3K27me3 level,39 are also noted in HCC, suggesting functional implication of these epigenetic regulators in HCC development. We further identified EZH2 and its associated PRC2 as one of the critical epigenetic regulators in HCC and demonstrated its tumor and metastasis promoting role in HCC development. Our findings are consistent with other previous reports on EZH2 up-regulation40 and tumorigenesis in HCC.41 Beyond this, our present findings provide new knowledge to understand how EZH2 contributes to HCC metastasis.

6C). Strikingly, EZH2-regulated miRNAs can potentially modulate cell motility-associated pathways and key signaling pathways. It was interesting to note that the first- and second-rated pathways were focal adhesion (Pathway ID hsa04510) and adherens junction (Pathway ID hsa04520), two crucial pathways in cancer cell invasion and metastasis. Consistent findings were obtained by PicTar, another miRNA target prediction algorithm (Supporting Fig. 5, Supporting Table 7). We also noticed that the RhoGTPase-associated cytoskeleton reorganization

axis was recurrently engaged RXDX-106 datasheet in six of the top-rated KEGG pathways, including those for focal adhesion (Pathway ID hsa04510), adherens junction (Pathway ID hsa04520), transforming growth factor

beta (TGF-β) signaling (Pathway ID hsa04350), noncanonical Wnt singling (Pathway ID hsa04310), axon guidance (Pathway ID hsa04360), and the actin cytoskeleton regulation (Pathway ID hsa04810) (Supporting Fig. 6). We previously reported that the RhoGTPase signaling pathway is frequently altered in human HCCs and is tightly associated with HCC metastasis.21, 35, 36 Our present findings further suggest that the EZH2-tumor suppressor miRNA axis may act upstream of the pathway to mediate its perturbation. Consistent with this notion, we found that knockdown of EZH2 resulted in down-regulation of RhoA and ROCK2 protein and inhibited stress fiber formation in HCC cells (Supporting Fig. 7). Taken together, the in silico analysis reinforces the tumor suppressive functions of EZH2-regulated miRNAs, and suggests their combinational effects in modulating key cell movement STI571 concentration and metastasis-related pathways in driving however HCC metastasis. Epigenetic regulation machinery involves multiple proteins with distinct functions. In our study, we first revealed that deregulation of epigenetic modifiers is common in HCCs. These epigenetic modifiers, including DNA methyltransferases, histone deacetylases, SET domain-containing histone methyltransferases, and

PcG proteins are direct mediators of epigenetic mechanisms. Their concordant deregulation reflects HCC epigenome is likely to be affected in multiple aspects. In line with our observation, not only are some of these proteins reported to be up-regulated in HCC,5, 37 but genome-wide DNA hypomethylation and promoter DNA hypermethylation of tumor-suppressors,38 as well as changes in global histone modification such as an increase of H3K27me3 level,39 are also noted in HCC, suggesting functional implication of these epigenetic regulators in HCC development. We further identified EZH2 and its associated PRC2 as one of the critical epigenetic regulators in HCC and demonstrated its tumor and metastasis promoting role in HCC development. Our findings are consistent with other previous reports on EZH2 up-regulation40 and tumorigenesis in HCC.41 Beyond this, our present findings provide new knowledge to understand how EZH2 contributes to HCC metastasis.

Thus, a large amount of well-characterized and highly qualified antibodies are needed in proteomics. Purified antigen, which is not always available, has proven to be one of the rate-limiting steps in mAb large-scale generation. find more Methods: Here we describe our strategies to establish a murine hybridoma cell bank for human liver plasma using unknown native proteins as the immunogens. The antibody-recognized plasma proteins were identified by MS following immunoprecipitation (IP), and by screening of human liver cDNA expression library. Results: We found that the established antibodies

reacted specifically with a number of important Candidate cancer biomarkers, Candidate cardiovascular disease genes include FN1 in plasma and extracellular matrix. The subcellular localization of these antigens was further confirmed by immune-histocytochemistry. 48 cases Different disease liver Pathological tissue specimens were also tested for their diagnostic value by 3 strain anti-FN1 antibodies and immune-histocytochemistry AZD3965 chemical structure diagnostic value. Conclusion: We believe these well-characterized antibodies would be useful in

diagnosis and treatment of Liver disease in the near future. Key Word(s): 1. FN1; 2. monoclonal antibody; 3. Liver disease; Presenting Author: JIAN WANG Additional Authors: PENGCHANG ZHU, FENWEI XIE Corresponding Author: FENWEI XIE Affiliations: Changzheng hospital Objective: FOXA2

functions as an important regulator in endoderm-derived organs development and body homeostasis. It has been reported that FOXA2 has a great impact on EMT process through targeting E-cadherin and Snail2. Additional, FOXA2 suppression is responsible for the TNF-α induced tumorigenesis in liver. However, the role of FOXA2 in HCC metastasis is still unknown. This study aims to clarify that FOXA2 functions as a suppressor of HCC metastasis. Methods: The expression of FOXA2 was tested in HCC patients’s Carbohydrate specimens including primary tumors, adjacent tissues and portal vein tumor thrombuses. The respectively FOXA2 expression and migration ability in different human hepatoma cell lines were also detected. The effects of overexpression and knockdown of FOXA2 were investigated in FOCUS and Hep-G2, respectively. We also evaluated the expression of FOXA2 in liver cancer tissue samples from 80 patients and analyzed the relationship between FOXA2 expression and clinicopathological features. Results: We demonstrated that expression of FOXA2 was down-regulated in the process of HCC development.

Thus, we cannot be certain that the rs-fc differences in this study are attributable to having CM. However, correlations between number of years with CM and atypical rs-fc are highly suggestive that our findings relate to the presence of CM. Because we did not have a cohort of episodic migraine subjects in this study, it is unclear if our findings are specific for CM or are applicable to episodic and CM. Migraine and control groups were not gender matched, potentially introducing a source of bias.[85] Also, subjects were not matched according to measures of anxiety and depression, conditions that may affect rs-fc between

pain regions. Considering the 3 functional connections differing between CM and controls that also correlated with the number of CM years, only one (anterior insula MLN8237 solubility dmso with PAG) also correlated with state anxiety scores. Eight CM subjects were using daily medications considered migraine prophylactic therapies (6 at doses considered sufficient for migraine prophylaxis). To explore the possibility that the use of these medications was driving our results, we performed post hoc analyses comparing rs-fc to the 5 pain ROIs in migraineurs taking prophylactic medications to migraineurs not taking prophylactic medications. There was no anatomic overlap between regions involved in the functional buy OSI-906 connections that differed between migraineurs and controls and regions involved in functional

connections that differed in migraineurs taking prophylactics and those not taking prophylactics. Thus, use of migraine prophylactic medications by a proportion of the migraineurs likely had little impact on our results reported herein. Also, CM subjects had a relatively short duration of CM (about 4 years). A longer duration of CM may be associated with more atypical rs-fc of pain regions. CM is associated with interictal atypical rs-fc of affective

pain regions with regions participating Nintedanib (BIBF 1120) in sensory-discriminative, cognitive, and integrative pain functions. Correlations between years with CM and the strength of some of these atypical functional connections suggest a causal relationship, although the direction of this relationship is uncertain. Atypical rs-fc of affective pain regions might relate to the abnormal affective processing of potentially painful stimuli and atypical affective responses to painful stimuli that are characteristic of CM. Studies comparing episodic migraine and CM and longitudinal studies are needed to determine if atypical rs-fc is a result of having CM or if atypical rs-fc predisposes the individual to developing CM. “
“To investigate if a headache frequency of 15 days per month constitutes a turning point in the psychosocial impairment associated with migraine. Migraine is differentiated into episodic and chronic forms based on a headache frequency criterion (< vs ≥15 headache days per month).

Bain – Advisory Committees or Review Panels: Astellas, Novartis, Merck, Astellas, Boehringer Ingelheim Darrell H. Crawford – Advisory Committees LDK378 in vitro or Review Panels: Roche Products Pty Ltd, Bristol Myers Squibb, Gilead Sciences, Novartis, MSD, Abbvie, Jansen; Consulting: Roche Products Pty Ltd; Grant/Research Support: Roche Products Pty Ltd; Speaking and Teaching: Roche Products Pty Ltd, Bristol Myers Squibb, Gilead Sciences, MSD Pietro Andreone – Advisory Committees or Review Panels: Roche, Janssen-Cilag, Gilead, MSD/Schering-Plough, Abbvie, Boehringer Ingelheim; Grant/Research Support: Roche, Gilead; Speaking and Teaching: Roche, MSD/Schering-Plough,

Gilead Tarek Hassanein – Advisory Committees or Review Panels: AbbVie, Bristol-Myers Squibb; Grant/Research Support: AbbVie Pharmaceuticals, Boehringer-Ingle-heim, Bristol-Myers Squibb, Eiasi Pharmaceuticals, Gilead Sciences, Janssen R&D, Idenix Pharmaceuticals, Ikaria Therapeutics, Merck Sharp & Dohme, Roche Pharmaceuticals, Ocera Therapeutics, Salix Pharmaceuticals, Sundise, TaiGen Biotechnology, Takeda Pharmaceuticals, Vital Therapies; Speaking and Teaching: Baxter, Bristol-Myers

Squibb, Gilead, Salix Wlodzimierz W. Mazur – Advisory Committees or Review Panels: Bristol-My-ers-Squibb company; Speaking and Teaching: Gilead, MSD, Roche, Abvee Sandra S. Lovell – Employment: AbbVie Barbara Da Silva-Tillmann – Employment: AbbVie Nancy Shulman – Employment: Abbvie Pazopanib concentration Massimo Puoti – Consulting: Abbvie Terry D. Box – Advisory Committees or Review Panels: Gilead, Genentech, Abb-Vie, Salix, Janssen; find more Grant/Research Support: Gilead, Merck, BMS, AbbVie, Idenix, Salix, Cumberland, Boehringer Ingelheim, Genfit, Vital Therapeutics, Sun-dise, Ikaria, Conatus; Speaking and Teaching: Gilead, Merck, Genentech, Salix Ira M. Jacobson – Consulting: Abbvie, Achillion, Boehringer Ingelheim, Bristol Myers Squibb, Gilead, Idenix, Genentech, Merck, Janssen,

Vertex; Grant/ Research Support: Abbvie, Boehringer Ingelheim, Bristol Myers Squibb, Gilead, Novartis, Genentech, Merck, Janssen, Vertex; Speaking and Teaching: Bristol Myers Squibb, Gilead, Genentech, Vertex, Janssen Introduction: In 2011-2012, approximately 30 patients were infected with genotype 1b HCV through nosocomial transmission during cardiac catheterization in a hospital in northern New England. Most of these patients were older and had cardiac comorbidities precluding treatment with interferon (IFN) and/or ribavirin (RBV). This study was conducted to offer IFN- and RBV-free treatment to these patients. Methods: Patients were enrolled and received open-label treatment with the fixed-dose combination of ledipasvir/sofosbuvir 90 mg/400 mg (LDV/SOF) for 12 weeks. In addition to efficacy and safety, T cell responses by ELISPOT and viral sequencing were assessed during and following treatment.

Ig). The recombinant plasmid was transfected into HEK293 cells using Lipofectamine 2000 (Invitrogen), and then VSIG4.Ig was purified from the culture supernatant using HiTrap Protein G HP Columns according to

the manufacturer’s recommendations (GE Healthcare). Mice were injected intravenously with either a lethal dose (25-30 mg/kg) or a sublethal dose (15 mg/kg) of ConA (Sigma-Aldrich). Serum alanine aminotransferase (ALT) levels were measured using a transaminase kit (Asan Pharmaceutical) according to the manufacturer’s Poziotinib chemical structure instructions. For adoptive transfer of KCs, KCs (3 × 106) isolated from VSIG4 WT or KO mice were injected intravenously into VSIG4 KO mice by way of the tail vein. Mouse livers were fixed in 4% paraformaldehyde, dehydrated, and embedded in paraffin. 5-μm sections were stained with hematoxylin and eosin using a standard procedure and analyzed by light microscopy. Liver MNCs were isolated by the collagenase digestion method with some modification.12–13 Briefly, mouse liver was perfused in situ with Hank’s buffered salt solution (HBSS) containing 0.025% collagenase, R788 in vitro removed, and passed through 70-μm stainless

steel mesh. Initial cell suspension that was resuspended in 40% Percoll was overlaid onto 70% Percoll and centrifuged at 750g for 20 minutes. MNCs were collected from the interface. For purification of KCs, liver MNC suspension was overlaid onto Percoll gradient (25%/50%), and centrifuged at 1,800g for 30 minutes. Megestrol Acetate KC-enriched MNCs located in the interface were harvested and stained with FITC-conjugated

anti-F4/80 (clone BM8, eBioscience). F4/80 positive KCs were purified using anti-FITC Microbeads (Miltenyi Biotech) according to the manufacturer’s protocols. KC isolates were 95% pure and KCs were the only cell fraction expressing VSIG4 among liver APCs (Supporting Fig. 1). For purification of splenic DCs, splenocytes were incubated with anti-CD11c Microbeads (Miltenyi Biotech) and enriched by the MACS system according to the manufacturer’s protocols. For purification of liver T- and NKT-cells, liver MNCs were stained with FITC-conjugated-NK1.1 mAb and PE Cy5-conjugated anti-TCR-β mAb, and then TCR-β+NK1.1+ NKT and TCR-β+NK1.1− T-cells were sorted using a BD FACSAria. T-cells (105) were plated in 96-well flat-bottom plates that were precoated with indicated concentrations of mouse anti-CD3e antibody (145-2C11) together with VSIG4.Ig or control Ig (10 μg/mL). [3H]-Thymidine (1 μCi/well) was added 16 hours prior to harvesting of the cultures. [3H]-Thymidine incorporation was measured with a Wallac MicroBeta TriLux Liquid Scintillation counter (PerkinElmer). In some experiments, purified DO11.10 T-cells (105) were incubated with KCs (1-10 × 103) in the presence of OVA323-339 (10 μg/mL) for 3 days before [3H]-thymidine incorporation. For liver NKT-cell tolerance induction, mice (Balb/c background) were injected intraperitoneally with α-GalCer (0.