7), suggesting that the aberration of either gene may be involved in the maintenance of aggressive phenotype of an established tumor. We also performed preliminary AZD9668 solubility dmso functional characterizations

of both putative drivers by siRNA-mediated target knockdown in HCC cell lines that carry the respective target amplification and compared with models without the amplification. We noted that results on BCL9 were mixed in the HUH6 cell line, which is copy number neutral with respect to BCL9, but had decreased viability upon BCL9 knockdown in one of the assays. Because BCL9 is involved in the Wnt/β-catenin–signaling pathway,[17] there may exist other mechanisms for activating this pathway in HUH6 cells: It has been shown that the Wnt pathway may be activated in the HUH6 cell line as a result of β-catenin mutations.[20] Blocking the Wnt/β-catenin pathway by knocking down BCL9 gene expression could then lead to tumor growth inhibition in HUH6 cells, which may be addicted Selleckchem VX 809 to this pathway for its tumorigenic properties. More research is needed to fully validate these two genes

as oncogenic drivers in HCC and to explore their utility in targeted cancer therapy. Our work nevertheless demonstrates a proof of concept that systematic clinical genomics approaches, such as the one presented here, could be valuable in uncovering novel, clinically relevant cancer driver genes, and that testing of such genes needs to be performed in relevant preclinical models, both with and without the corresponding genetic aberration. Future directions of our work include high-throughput dropout screens to systematically test all genes within the focal amplicons, an unbiased approach similar to the forward genetic screening by Sawey et al.[9] One of the biggest challenges in CNA-driven target identification is to distinguish true driver gene(s) from

passengers in a focal amplicon. It has been shown that multiple drivers may even coexist in a highly focal amplicon, such as CCND1 and FGF19.[9] It would be valuable to perform unbiased screening to validate all candidate somatic CNA drivers selleck inhibitor in appropriate models and then dissect key attributes that distinguish drivers from passengers to facilitate future in silico algorithm development. Toward this end, the genomic characterization of a comprehensive collection of 30 HCC cell line models in our study will also serve as a valuable resource for future research in this direction. The authors thank Drs. John Lamb and Soonmyung Paik for scientific discussion in this study, Peter C. Roberts for facilitating data management and transfer, and Sylvie Sakata for study support. Additional Supporting Information may be found in the online version of this article. Figure S1. Association between somatic CNA, mRNA expression and clinical outcome.

After a 1 g/kg dose of fructose, blood levels increase minimally to just ∼0.5 mM,22 much less than the 10 mM increase found with an equivalent dose of glucose. Fructose metabolism also differs from glucose metabolism in that uptake is relatively unregulated by insulin.25 Fructokinase action is 10 times faster than glucokinase and hexokinase, and fructose accumulates

in the liver as fructose-1-phosphate.26 Perfusion studies of liver tissue show that this step is rapid enough to precipitate a depletion of adenosine triphosphate (ATP) content to 23%, although ATP recovers to normal within 40 minutes.27 Fructose-1-phosphate is converted into triose phosphates, which become substrates for gluconeogenesis or the downstream Venetoclax concentration steps of glycolysis and DNL. In a 6-hour study tracking the fate of an oral bolus of labeled fructose, 35% of fructose was oxidized, 0.4% appeared as FFA in newly formed VLDL-TG, 38% appeared as glycerol

in VLDL-TG, and some remained unaccounted for, likely remaining in the liver in the HSP inhibitor review form of glycogen.28 In sum, fructose metabolism is unique from glucose; it enters the liver in a relatively unregulated fashion and is metabolized into products of both glycolysis and gluconeogenesis.29 Paradoxically, although fructose does not increase insulin acutely, over time it increases insulin resistance, fasting glucose, and insulin. Dirlewanger et al.30 found that fructose induces hepatic and extrahepatic insulin resistance in healthy adult humans in infusion/clamp studies, although the mechanism of how insulin resistance is induced is not known. High fructose consumption clearly increases visceral fat in healthy adults and in animal models (see Supporting Material). In selleck screening library a 10-week study, subjects consuming fructose beverages gained significantly more visceral adiposity compared

to those consuming eucaloric glucose beverages.31 A cross-sectional study of adolescents also found a relationship between high fructose consumption and visceral adiposity.32 It may be that induction of visceral fat results in increased insulin resistance because visceral fat is thought to be inherently “diabetogenic.”33 However, it is also possible that the deposition of lipids in the liver causes insulin resistance and leads to increased visceral adiposity.33 Stanhope and Havel34 postulate that decreased insulin stimulation by fructose leads to decreased lipoprotein lipase activity in saturated adipose tissue and increased lipoprotein lipase activity in visceral adipose tissue, thus leading to increased lipid uptake into the hypertrophied adipocytes. In 1970, Mann et al.35 demonstrated that sucrose reduction in the diet resulted in improved TG levels in healthy men. This finding continues to be supported by numerous studies demonstrating a hypertriglyceridemic effect of fructose in humans.

19, 0.80-1.78) was not.

Conclusions: Younger (<40y) HCV LT recipients have significantly reduced graft survival, higher rates of re-LT and HCV-related death than older HCV recipients. This suggests a more aggressive natural history for HCV disease post-LT and identifies a group in need of early consideration of HCV therapy. Disclosures: Norah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Consulting: BMS, Merck; Grant/Research Support: Eisai, Biotest, Vertex, Gilead, AbbVie, Novartis, Merck The following people have nothing to disclose: Varun Saxena, Jennifer L. Dodge, John P. Roberts Background/Aims: To identify the impact of portal vein thrombosis (PVT) on post liver transplant Opaganib (LT) outcomes along with other covariates and assess factors associated with complications amongst PVT patients. Methods: ABT-263 order Retrospective cohort study of 621 adult LT recipients (University of Alberta, London Health Sciences Centre) between 01/2002-12/2012. PVT

was identified in 147 (24%) patients and 474 (76%) non-PVT patients served as controls. Cox survival analysis was performed to determine independent associations with overall mortality. Results: Demographic factors (mean age 53, 69% male) were similar between groups. There were also no differences in mean MELD (PVT 19 vs. controls 19, p=0.9) and Child Pugh scores (10 vs. 10, p=0.9) on the day of LT. Donor factors (mean DRI:1.6 vs. 1.5, p=0.2) were similar. Using Cox multivariable survival analysis, covariates independently associated with overall mortality included Age (adjusted Hazard ratio ∼ aHR 1.02, p=0.015) and requiring ICU support pre-LT (aHR 2.17, p=0.006), but not PVT (p=0.67). 5-year survival was similar between PVT and controls (75%,p= 0.8). In comparing PVT patients who did not survive (n=32) with PVT survivors (n=115), non-survivors (n=32) were more likely to have complete thrombus occlusion (38% vs. 13%, p=0.027) and

hepatofugal flow (31% vs. 13%, p=0.08). Non-survivors were more likely require thrombectomy (69 vs. 31%, p=0.08) and develop reocclusion post-LT (16% vs. 3%, p=0.024). Anti-coagulation rates were similar between groups. Conclusion: Well-selected LT patients who had PVT prior to LT have similar post-LT outcomes with selleck controls when adjusting for donor and recipient factors. Subgroups of PVT LT patients who did worse post-LT (complete thrombosis pre-LT, thrombectomy at LT and reocclusion post-LT) warrant closer evaluation in listing and management post-LT. Adjusted survival (Cox) for PVT LT recipients vs. controls (p=0.67). Disclosures: Constantine J. Karvellas – Grant/Research Support: Merck; Speaking and Teaching: Gambro The following people have nothing to disclose: Filipe S. Cardoso, Malcolm M. Wells, Fayaz A. Handoo, Lukasz Kwapisz, Mansour G. Alghanem, Norman Kneteman, Paul Marotta, Bandar Al-Judaibi Background.

19, 0.80-1.78) was not.

Conclusions: Younger (<40y) HCV LT recipients have significantly reduced graft survival, higher rates of re-LT and HCV-related death than older HCV recipients. This suggests a more aggressive natural history for HCV disease post-LT and identifies a group in need of early consideration of HCV therapy. Disclosures: Norah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Consulting: BMS, Merck; Grant/Research Support: Eisai, Biotest, Vertex, Gilead, AbbVie, Novartis, Merck The following people have nothing to disclose: Varun Saxena, Jennifer L. Dodge, John P. Roberts Background/Aims: To identify the impact of portal vein thrombosis (PVT) on post liver transplant PF-02341066 cost (LT) outcomes along with other covariates and assess factors associated with complications amongst PVT patients. Methods: 3-deazaneplanocin A order Retrospective cohort study of 621 adult LT recipients (University of Alberta, London Health Sciences Centre) between 01/2002-12/2012. PVT

was identified in 147 (24%) patients and 474 (76%) non-PVT patients served as controls. Cox survival analysis was performed to determine independent associations with overall mortality. Results: Demographic factors (mean age 53, 69% male) were similar between groups. There were also no differences in mean MELD (PVT 19 vs. controls 19, p=0.9) and Child Pugh scores (10 vs. 10, p=0.9) on the day of LT. Donor factors (mean DRI:1.6 vs. 1.5, p=0.2) were similar. Using Cox multivariable survival analysis, covariates independently associated with overall mortality included Age (adjusted Hazard ratio ∼ aHR 1.02, p=0.015) and requiring ICU support pre-LT (aHR 2.17, p=0.006), but not PVT (p=0.67). 5-year survival was similar between PVT and controls (75%,p= 0.8). In comparing PVT patients who did not survive (n=32) with PVT survivors (n=115), non-survivors (n=32) were more likely to have complete thrombus occlusion (38% vs. 13%, p=0.027) and

hepatofugal flow (31% vs. 13%, p=0.08). Non-survivors were more likely require thrombectomy (69 vs. 31%, p=0.08) and develop reocclusion post-LT (16% vs. 3%, p=0.024). Anti-coagulation rates were similar between groups. Conclusion: Well-selected LT patients who had PVT prior to LT have similar post-LT outcomes with selleck controls when adjusting for donor and recipient factors. Subgroups of PVT LT patients who did worse post-LT (complete thrombosis pre-LT, thrombectomy at LT and reocclusion post-LT) warrant closer evaluation in listing and management post-LT. Adjusted survival (Cox) for PVT LT recipients vs. controls (p=0.67). Disclosures: Constantine J. Karvellas – Grant/Research Support: Merck; Speaking and Teaching: Gambro The following people have nothing to disclose: Filipe S. Cardoso, Malcolm M. Wells, Fayaz A. Handoo, Lukasz Kwapisz, Mansour G. Alghanem, Norman Kneteman, Paul Marotta, Bandar Al-Judaibi Background.

“
“Background and Aim:  MicroRNAs are short noncoding RNA molecules that are responsible for the posttranscriptional regulation of target genes. The aim of this study was to determine whether microRNA-199b-5p (miR-199b) plays a role find more in the progression and prognosis of hepatocellular carcinoma (HCC), and to elucidate whether hypoxia-inducible factor-1α (Hif1α) is regulated by miR-199b. Methods:  In this study, 35 matched

HCCs and cirrhosis tissues were assayed for miR-199b and Hif1α expression. To evaluate the role of miR-199b, we assessed cell proliferation rate and clonogenic survival of miR-199b- or negative control-transfected cells by MTT and clone formation assay, respectively. In addition, the regulation of Hif1α by miR-199b was evaluated by Western blotting and luciferase assay. MiR-199b was downregulated in 77% of HCCs, whereas Hif1α protein was upregulated in 69% of cases. A significant inverse correlation between miR-199b and Hif1α was observed in HCCs. Results:  Patients with lower levels of miR-199b expression had poorer overall survival and progression-free survival

rates, whereas patients with higher levels of miR-199b expression had better survival. Moreover, miR-199b could restrain cell growth and obviously enhance the radiosensitizing effect of HepG2 cells. MiR-199b and pGL3-Hif1α vector-transfected cells showed suppressed Hif1α protein expression and significant reduced luciferase activity. Conclusions:  Underexpressed miR-199b, which may be via the upregulation of Hif1α in HCCs, is inversely correlated with survival and directly correlated with the malignant status DAPT of HCC patients. “
“Partial hepatectomy (PH) induces robust hepatic regenerative and metabolic responses that are considered to be triggered by humoral factors. see more The aim of the study

was to identify plasma protein factors that potentially trigger or reflect the body’s immediate-early responses to liver mass reduction. Male C57BL/6 mice were subjected to sham operation, 70% PH or 90% PH. Blood was collected from the inferior vena cava at 20, 60 and 180 min after surgery. Using a label-free quantitative mass spectrometry-based proteomics approach, we identified 399 proteins exhibiting significant changes in plasma expression between any two groups. Of the 399 proteins, 167 proteins had multiple unique sequences and high peptide ID confidence (>90%) and were defined as priority 1 proteins. A group of plasma proteins largely associated with metabolism is enriched after 70% PH. Among the plasma proteins that respond to 90% PH are a dominant group of proteins that are also associated with metabolism and one known cytokine (platelet factor 4). Ninety percent PH and 70% PH induces similar changes in plasma protein profile. Our findings enable us to gain insight into the immediate-early response of plasma proteins to liver mass loss.

This experimental approach provides quantitative and mechanistic data regarding the role of specific oncogenes

during hepatocarcinogenesis. BrdU, bromodeoxuridine; CHeGA, comparative hepatocyte growth assay; EO, extreme outlier; hPAP, human placental alkaline phosphatase; lacZ, Selleckchem Opaganib β-galactosidase; TAg, simian virus 40 T antigen; TGFα, transforming growth factor alpha; uPA, urokinase-type plasminogen activator. Mice were housed and maintained according to The Guide for the Care and Use of Laboratory Animals in Association for Assessment and Accreditation of Laboratory Animal Care–accredited facilities. All experimental procedures were approved by the Institutional Animal Care and Use Committee. The transgenic lines used in selleck these studies have been assigned the following genetic designations: major urinary protein uPA line 350-2, TgN(MupPlau)1Eps; hsMT-nLacZ line 379-4, TgN (Mt1nLacZ)4Eps; R26-hPAP line 808-6, TgN(R26ALPP)5Eps; AL-TAg line 522-8, TgN(Alb1SV)46Bri; MT-TGFα line 1745-8, TgN(Mt1Tgfa)149Bri;

AL-c-myc line 741-3, TgN(Alb1Myc)82Bri.3, 5, 14 For these studies, mice were of the FVB, C57BL/6, or (FVB6)F1 strain background. One group of recipient mice were athymic Swiss nu/nu. Transplant recipient mice carrying metallothionein (MT)-TGFα donor hepatocytes were administered 25 mM zinc sulfate in drinking water starting at the time of transplant to induce transgene expression.5 β-Galactosidase (LacZ)-marked or human placental alkaline phosphatase (hPAP)-marked donor hepatocytes were isolated from 2-week-old to 5-week-old donor mice using a modified two-step ethylenediaminetetra-acetic acid/collagenase A protocol.14 In all cases, mice were excluded as donors if they displayed focal lesions visible on gross examination. Transgenic mouse livers lacking see more these alterations contain few or no areas of parenchyma that would be microscopically

diagnosed as neoplastic, although dysplastic cells may be present.3, 5, 6, 12 The concentration of viable large cells (hepatocytes) was determined by trypan blue exclusion using a hemacytometer. Cells were maintained at 4°C until transplanted. Hepatocytes were transplanted surgically in 10 μl of L15 medium (Life Technologies, Rockville, MD) via intrasplenic injection into histocompatible recipients within 6 hours of isolation.14 Recipient mice were administered 0.1 mg/kg cadmium sulfate intraperitoneally to induce expression of the MT-nLacZ transgene, then 16 to 24 hours later liver was collected and a portion was fixed in 4% paraformaldehyde at 4°C for 1 hour then transferred to 70% ethanol. β-Galactosidase- and hPAP-expressing hepatocyte foci were identified as described,14 by incubating separate pieces of liver with an appropriate enzyme-selective substrate. Transgene-expressing cells displayed a blue reaction product.

01); Treatment with UA, data were presented as relative reduce compared with leptin treatment (all P < 0.01). HSC-T6 were treated for 30 minutes with Leptin, the PI3K, p-Akt, p-P38MAPK levels were distinctly increased compared with Selleckchem Hydroxychloroquine normal control group

(all P < 0.01); Treatment with UA, data were presented as relative reduce compared with leptin treatment (all P < 0.01). HSC-T6 were treated for 24 hours with Leptin, TIMP-1 level was increased compared with normal control group (P < 0.05); Treatment with UA, data were presented as obvious reduce compared with leptin treatment (P < 0.01); while MMP-1 level was decreased compared with normal control group (P < 0.05); Treatment with UA, data were presented as obvious increase compared with leptin treatment (P < 0.01). Conclusion: UA decreases the proteins expression of NOX subunit gp91phox, p22phox, p67phox, Rac1, PI3K, p-Akt and p-p38MAPK induced by leptin in Rat HSC-T6. UA can decrease protein expression of TIMP-1 and increase MMP-1 expression. The mechanism may be related to inhibiting activation of NOX-PI3K/Akt and P38MAPK signal net. Key Word(s): 1. HSCs; 2. ursolic acid; 3. NOX oxidase; 4. PI3K/Akt and P38MAPK; Presenting

Author: LU CHEN Additional Authors: WENHUA HE, FENG SHI, WEN HUANG, TAO CHEN, DEQIANG HUANG, XUAN ZHU Corresponding Author: XUAN ZHU N Affiliations: Nanchang University Objective: To explore the mechanism and effects this website of UA on hedgehog (Hh) signal pathway YAP-TEAD Inhibitor 1 cost in hepatic stellate cell (HSC-T6). Methods: HSC-T6 in the exponential growth phase were devided into four groups: normal control group; leptin (100 ng/ml) treated; UA (50 μM) treated; DPI (20 μM) treated; leptin treated together with UA (50 μM) and leptin treated together with NOX inhibitor DPI (20 μM). HSC-T6 were treated with medicine for 12 hours and mRNA expression of Shh, smo, Gli1/2 were analyzed with RT-PCR. HSC-T6 were treated with medicine for 24 hours and protein expression of Gli2

and Rac1 were analyzed with Western blotting. HSC-T6 were treated with medicine for 12 hours, 24 hours, 48 hours and HSC-T6 proliferation was analyzed with MTT. Results: HSC-T6 were treated for 12 hours with Leptin, UA and DPI both decrease the mRNA expression of Shh, Smo, Gli2 induced by leptin (all P < 0.01), but leptin, UA and DPI had no effect on the mRNA expression of Gli1(P > 0.05). HSC-T6 were treated for 24 hours with Leptin, UA and DPI both decreases the protein expression of Rac1, Gli2 induced by leptin (all P < 0.01). HSC-T6 were treated for 12 hours with Leptin, leptin promote the HSC-T6 proliferation (P < 0.01), and UA can inhibits the HSC-T6 proliferation induced by leptin (P < 0.01). Conclusion: UA can inhibit expression of Shh, Smo, Gli2 mRNA and lower expression of Gli2 protein in hedgehog signal pathway of HSC-T6 induced by the leptin, and inhibit HSC-T6 growth and proliferation.

01); Treatment with UA, data were presented as relative reduce compared with leptin treatment (all P < 0.01). HSC-T6 were treated for 30 minutes with Leptin, the PI3K, p-Akt, p-P38MAPK levels were distinctly increased compared with http://www.selleckchem.com/products/nu7441.html normal control group

(all P < 0.01); Treatment with UA, data were presented as relative reduce compared with leptin treatment (all P < 0.01). HSC-T6 were treated for 24 hours with Leptin, TIMP-1 level was increased compared with normal control group (P < 0.05); Treatment with UA, data were presented as obvious reduce compared with leptin treatment (P < 0.01); while MMP-1 level was decreased compared with normal control group (P < 0.05); Treatment with UA, data were presented as obvious increase compared with leptin treatment (P < 0.01). Conclusion: UA decreases the proteins expression of NOX subunit gp91phox, p22phox, p67phox, Rac1, PI3K, p-Akt and p-p38MAPK induced by leptin in Rat HSC-T6. UA can decrease protein expression of TIMP-1 and increase MMP-1 expression. The mechanism may be related to inhibiting activation of NOX-PI3K/Akt and P38MAPK signal net. Key Word(s): 1. HSCs; 2. ursolic acid; 3. NOX oxidase; 4. PI3K/Akt and P38MAPK; Presenting

Author: LU CHEN Additional Authors: WENHUA HE, FENG SHI, WEN HUANG, TAO CHEN, DEQIANG HUANG, XUAN ZHU Corresponding Author: XUAN ZHU N Affiliations: Nanchang University Objective: To explore the mechanism and effects selleck chemicals llc of UA on hedgehog (Hh) signal pathway CX-5461 price in hepatic stellate cell (HSC-T6). Methods: HSC-T6 in the exponential growth phase were devided into four groups: normal control group; leptin (100 ng/ml) treated; UA (50 μM) treated; DPI (20 μM) treated; leptin treated together with UA (50 μM) and leptin treated together with NOX inhibitor DPI (20 μM). HSC-T6 were treated with medicine for 12 hours and mRNA expression of Shh, smo, Gli1/2 were analyzed with RT-PCR. HSC-T6 were treated with medicine for 24 hours and protein expression of Gli2

and Rac1 were analyzed with Western blotting. HSC-T6 were treated with medicine for 12 hours, 24 hours, 48 hours and HSC-T6 proliferation was analyzed with MTT. Results: HSC-T6 were treated for 12 hours with Leptin, UA and DPI both decrease the mRNA expression of Shh, Smo, Gli2 induced by leptin (all P < 0.01), but leptin, UA and DPI had no effect on the mRNA expression of Gli1(P > 0.05). HSC-T6 were treated for 24 hours with Leptin, UA and DPI both decreases the protein expression of Rac1, Gli2 induced by leptin (all P < 0.01). HSC-T6 were treated for 12 hours with Leptin, leptin promote the HSC-T6 proliferation (P < 0.01), and UA can inhibits the HSC-T6 proliferation induced by leptin (P < 0.01). Conclusion: UA can inhibit expression of Shh, Smo, Gli2 mRNA and lower expression of Gli2 protein in hedgehog signal pathway of HSC-T6 induced by the leptin, and inhibit HSC-T6 growth and proliferation.

Pseudocontinuous ASL was collected using 30 pairs of tag and control acquisition using a 3-dimensional gradient-echo spin-echo (GRASE) acquisition. All images were registered to a high-resolution anatomical atlas. Average CBF measurements

within regions of contrast-enhancement and T2 hyperintensity were evaluated between the two modalities. Additionally, voxel-wise correlation between CBF measurements obtained with DSC and ASL were assessed. Results demonstrated a positive linear correlation between DSC and ASL measurements of CBF when regional average values were compared; however, selleck a statistically significant voxel-wise correlation was only observed in around 30-40% of patients. These results suggest DSC and ASL may provide regionally similar, but spatially different measurements of

CBF. Magnetic resonance imaging (MRI) is the mainstay of brain tumor imaging, both in diagnosis and treatment. Traditionally, clinicians rely on contrast enhancement to characterize the relative degree of malignancy in suptratentorial BMS-777607 tumors. However, with increasing evidence for the critical role of angiogenesis in determination of tumor malignancy and growth potential, imaging modalities capable of quantifying cerebral blood flow (CBF) have become attractive alternatives. Several studies have shown that higher grade brain tumors have significantly higher perfusion measurements than low-grade tumors,[1-3] suggesting that CBF measurements may be a better method

for characterizing brain tumor angiogenesis and monitor treatment response. As antiangiogenic therapy is now the standard of care for recurrent malignant gliomas, there is a significant need for monitoring changes in cerebral blood flow within selleck chemicals llc areas of suspected tumor independent of contrast enhancement. The gold standard for perfusion MR imaging is dynamic susceptibility contrast (DSC) MRI, which uses a bolus injection of paramagnetic contrast agent, usually gadolinium, as a nondiffusible tracer for CBF. Calculations of CBF, CBV (cerebral blood volume; the fraction of tissue volume occupied by blood), and mean transit time (MTT = CBF/CBV, the time it takes for blood to pass through the vasculature within the tissue of interest) can be made simultaneously. However, this requires deconvolving the arterial input function (AIF) from the time series data. As a result, few studies have been done on the reproducibility of DSC measurements of CBF. Arterial spin labeling (ASL) is a continually evolving noninvasive technique for quantifying CBF. ASL uses magnetically tagged blood water as an endogenous, diffusible tracer for blood flow. Specifically, blood in a feeding artery is subjected to an inversion pulse, and the magnetization can be followed as it is transferred to brain tissue by capillary exchange at a rate dependent on perfusion of the tissue.

Similarly, the number

of BrdU+CD8β+ cells, BrdU+FoxP3+ cells, or total FoxP3+ cells in the portal area were counted. All counting was performed in a blinded fashion. The phenotypes of donor MHCII+ cluster-forming cells were analyzed, as reported previously,6 in fresh serial 2-μm cryosections of the parathymic LNs and graft liver. DCs in the liver nonparenchymal cells and hepatic lymph were defined as the MHCIIhighCD103high population, based on fluorescence-activated cell-sorting (FACS) data (Fig. 1A,D) and in accord with earlier studies.3, 12 Those DCs in the healthy MEK inhibitor rat liver could be subdivided into three phenotypically different groups: CD172a+CD11b+ DCs (∼44%); CD172a+CD11b− DCs (∼20%); and CD172a−CD11b+ DCs (∼28%) (Fig. 1A,B). Five days after sublethal irradiation, the total number of liver DCs decreased from ∼2.8 × 105 to ∼5.1 × 104 (Fig. 1C), and the percentages of the three subsets changed to ∼64%, ∼28%, and ∼5%, respectively (Fig. 1A,B). Notably, the CD172a−CD11b+ subset was radiosensitive and decreased dramatically after irradiation, but ∼25% of the other two subsets remained. MHCIIhighCD103high DCs in the Decitabine chemical structure hepatic lymph could also be subdivided into three phenotypically different groups: CD172a+CD11b+ (∼80%); CD172a+CD11b− (∼15%); and CD172a−CD11b+ DCs (∼3%) (Fig. 1D,E). Five days after

sublethal irradiation, the total number of lymph DCs decreased from ∼1.3 × 105 to ∼1.4 × 104 (Fig. 1F), and the percentages of the three subsets changed to ∼90%, ∼8%, and ∼1%, respectively

(Fig. 1D,E). Thus, among lymph DCs, the CD172a+CD11b+ subset was relatively radioresistant, with ∼13% remaining after irradiation, whereas the other two subsets were very radiosensitive and were almost abolished. As in our previous study,6 donor MHCII+ and donor MHCI+ cells readily migrated to the recipient’s secondary lymphoid organs (i.e., the spleen, skin LNs, this website and Peyer’s patches), and donor MHCII+ DCs formed clusters with recipient BrdU+ cells in the T-cell areas on days 1-3 after LT in the Irr(−) group (Supporting Fig. 1A,C). Because Peyer’s patches do not possess afferent lymphatics, DC entry should be through the blood, presumably through the high endothelial venules.6 FACS analysis of skin LNs (Fig. 2A) and Peyer’s patches (not shown) revealed that more than 90% of the migrated donor MHCIIhighCD103high DCs were CD172a+CD11b−. The exception was the parathymic LNs, in which both CD172a+CD11b− and CD172a+CD11b+ donor DCs were found (Fig. 2A); the CD172a+CD11b+ subset constituted ∼70% of all DCs. In the Irr(−) group, donor MHCII+ and MHCI+ cells appeared in the peritoneal cavity on days 1-3 after LT. There were comparable numbers of donor cells in the Irr(+) group (Fig. 3A).