15, 16 Additionally, the expression of NK cell inhibitory ligand MHC I is down-regulated,17 whereas TRAIL receptor expression is up-regulated in activated HSCs,18, 19 which may represent additional important mechanisms contributing to increased sensitivity

of HCSs to NK cell/TRAIL killing. At present, it is not clear whether expression of NKG2D ligands (MICA/B) is also up-regulated in activated human HSCs during chronic liver diseases. Although Kahraman et al.13 performed immunohistochemical analyses of NASH liver samples with MICA/B antibodies, the expression of MICA/B on the HSCs LY294002 was not well illustrated. Double-staining with MICA/B antibodies and HSC markers will be required in the future to identify whether MICA/B proteins are expressed on activated HSCs in patients with chronic liver disease. In addition to having

a beneficial effect on liver fibrosis, the interaction of NKG2D and corresponding ligands may also play an important role in immunosurveillance against cholangiocarcinoma and hepatocellular carcinoma. Genetic analyses of NKG2D polymorphisms strongly suggest that interaction of NKG2D-MICA protects against cholangiocarcinoma in patients with primary sclerosing cholangitis.20 The inhibitory effect of NKG2D on liver tumors is likely mediated via activation of NK cells and the subsequent production of TRAIL, which is a potent cytotoxic mediator that kills hepatocellular carcinoma and cholangiocarcinoma cells.21 However, MICA/B proteins are also cleaved proteolytically from hepatocellular carcinoma cells GDC-0449 molecular weight and appear as soluble particles in serum from these patients.22 These soluble MICA/B proteins can down-regulate NKG2D expression and inhibit NKG2D-mediated NK cytotoxicity against liver tumor cells,23 allowing tumor cells to escape from immunity attacks mediated

by NKG2D. In summary, interaction between NKG2D and ligands can trigger NK cell activation, playing an important role in host defenses against hepatitis viral infection and liver cancer development, as well as the inhibition of liver fibrosis (Fig 1). Such activation may also contribute to hepatocellular damage in patients with HCV infection and NAFL/NASH. Additionally, the down-regulation MCE of this activation (e.g., by alcohol drinking) may accelerate the progression of liver disease, including viral infection and liver fibrosis,24, 25 while up-regulation of NKG2D expression on NK cells by poly I:C and IFN-γ can markedly enhance NK cell cytotoxicity against hepatocytes and HSCs, leading to hepatocellular damage and the reduction of liver fibrosis, respectively.11, 15 Interestingly, treating mice with IFN-α also markedly increased expression of NKG2D on liver NK cells (our unpublished data).

19 × 10−3 (αcorrect = 2.38 × 10−3/2). Spearman’s r test was used to analyze the correlation between XRCC4 genotypes and XRCC4 protein expression. Kaplan-Meier’s survival analysis with the log-rank test was used to evaluate the relationship between this polymorphism and HCC Selleck Copanlisib prognosis. Risk factors for HCC prognosis were first selected using Cox’s multivariate regression model (including age, sex, race, HBV and HCV infection, AFB1 exposure information, tumor size, tumor TNM stage, and all possible multiplicatively interactive variables)

with step-wise forward selection based on the likelihood ratio test. Hazard ratios (HRs) and 95% CIs for risk factors were next calculated in the same multivariate Cox’s regression model (simultaneously including all risk variables and multiplicatively interactive variables). Our final analysis included 1,499 HCC cases and 2,045 controls (Supporting Table 3). There were no significant differences between cases and controls in terms of distribution of age, sex, race, and HBV and HCV status as a result of individual matching (P > 0.05). These results suggest that HCC patient data were comparable to control data. The AFB1 exposure information for the entire study population is shown in Supporting BMS-354825 solubility dmso Table 4. We observed that the AFB1 exposure years were associated with

an increased risk for HCC (OR = 2.53 for medium-exposure years; OR = 5.85 for long-exposure years). We also found that MCE HCC cases (28.38 fmol/mg) had higher serum levels of AFB1 ALB adducts than controls (11.57 fmol/mg).

For statistical analysis, values were logarithmically transformed and then were divided into three stratus: low (<2.18 ln fmol/mg), medium (2.18-2.98 ln fmol/mg), and high (>2.98 ln fmol/mg), according to the mean logit value of serum AFB1 ALB adducts among controls and cases (Supporting Table 4). Regression analysis showed that HCC risk gradually increased with an increasing number of exposure levels (adjusted OR = 2.10-6.52; P < 0.01). XRCC4 genotypes from all peripheral blood DNA samples are listed in Table 1. For these SNPs, the genotype distribution of controls was consistent with those expected from Hardy-Weinberg's equilibrium (Supporting Table 5; P > 0.05). Among 21 SNPs, only rs28383151 (no. 3) was observed to modify HCC risk. Logistic regression analyses exhibited that the adjusted OR for HCC for those individuals carrying the heterozygotes of rs28383151 G and A allele (rs28383151-GA) versus those with the homozygotes of the rs28383151 G alleles (rs28383151-GG) was 1.94 (95% CI: 1.54-2.43); the corresponding OR for those featuring the homozygotes of rs28383151 A alleles (rs28383151-AA) was 3.10 (95% CI: 2.10-4.58).

We also identified TGFβ2 as a good candidate biomarker for ICC prognosis. Importantly, osteopontin selleck inhibitor and TGFβ2 protein expression were the most correlated independent variables (Supporting Table 5). Accordingly, differences in OS (P = 0.06) and DFS (P = 0.008) could be also observed by combining the expression of TGFβ2 and osteopontin (Supporting Fig. 3). Identifying ICC with favorable or unfavorable prognoses might orientate the selection of the most appropriate treatment, including liver resection/transplantation, chemotherapy, and targeted therapy,

either alone or in combination. Although ICC is not currently a widely accepted indication for orthotopic liver transplantation (OLT), some studies suggest that OLT could be indicated for selected ICC patients, as suggested for hilar cholangiocarcinoma.[47] A combination of neoadjuvant ABT-263 chemical structure therapy followed by OLT in appropriately selected patients with unresectable ICC also demonstrated promising disease recurrence-free survival.[48] Given that the expression of osteopontin correlates with relevant clinical variables (OS, DFS, hilar lymph nodes, macrovascular invasion), we believe that patients with

low to absent expression may benefit from OLT. In conclusion, by using an unsupervised approach we showed a clear correlation between genomic changes in the stroma and the aggressiveness of ICC, and we identified osteopontin as a promising prognostic biomarker. In addition, these observations support the idea that targeting the tumor stroma may represent a valid and innovative therapeutic strategy in ICC. The authors thank the Plateforme Génomique Santé, the Centre de Ressources Biologiques Santé (Rennes), the

Liver Biobanks Network, and Pascale Bellaud from the H2P2 histopathological platform (Biosit, Rennes). C.C. thanks Dr. Wendy T. Watford from the University of Georgia 上海皓元医药股份有限公司 for critically reviewing the article. Additional Supporting Information may be found in the online version of this article. “
“Background and Aim:  Aplasia ras homolog member I (ARHI) is a maternally imprinted tumor suppressor gene. ARHI protein is widely expressed in many types of human tissues; however, its expression is frequently reduced or absent in various tumors and plays a tumor suppressor role for in vitro study. In this study, we investigated the expression level of ARHI in gastric cancer in order to investigate the function of ARHI and signaling pathways that might be linked during gastric cancer development. Methods:  ARHI mRNA and protein expression levels were analyzed in primary gastric cancer tissues, adjacent noncancerous gastric tissues and gastric cancer cell lines using semi-quantitative polymerase chain reaction, western blotting and immunohistochemistry, respectively. Results:  Our results showed that both mRNA and protein expression levels of the ARHI gene were significantly downregulated (P < 0.

Natural products continue to be an invaluable source for anticancer drug discovery. In recent years the natural omega-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA) has been shown to possess

promising anticancer properties. Currently, dietary consumption remains the only means of acquiring DHA. With this form of intake DHA’s activity is diluted and markedly reduced as it is incorporated into plasma phospholipids or proteins. Clearly if the anticancer potential of this natural lipid is to be fully realized, novel delivery strategies must be employed. Herein, we evaluate in an in vitro cell culture system the utility of the low-density lipoprotein (LDL) as a nanoscale delivery vehicle for DHA. Methods: LDL-DHA nanoparticles were prepared and subject to extensively biophysical characterization. The therapeutic utility of LDL-DHA was evaluated in normal and malignant murine hepatocyte Epacadostat datasheet cell lines, TIB-73 and TIB-75 respectively, using MTT dose response, www.selleckchem.com/products/Trichostatin-A.html FACS, confocal microscopy and oxidative stress analyses. Results: Engineered LDL nanoparticles, uniformly loaded with unesterified DHA (LDL-DHA), closely resembled native LDL morphologically and biochemically. With regards to its biological activity,

LDL-DHA nanoparticles were avidly taken up by both TIB-73 and TIB-75 cells. Dose response evaluations revealed that LDL-DHA was selectively cytotoxic to the malignant TIB-75 cells. The selectivity of LDL-DHA was further exemplified with co-cultures of these two cells, therapeutic doses of LDL-DHA that completely killed the TIB-75 proved to be innocuous to TIB-73 leaving them unharmed. FACS analysis

showed that LDL-DHA activated both apoptotic and necrotic death pathways in the TIB-75 cells. Additional studies went on to show that LDL-DHA treatment selectively induced pronounced lipid peroxidation and oxidative stress in malignant TIB-75 cells. These pathways play a central role in LDL-DHA mediated cancer cell kill as supplementation medchemexpress with vitamin E was able to rescue the TIB-75 cells. Conclusion: These studies collectively demonstrate that LDL-DHA nanoparticles shows great promise as a selective anticancer agent against hepatocellular carcinoma. Disclosures: The following people have nothing to disclose: Ian Corbin, Lacy Reynolds, Rohit Mulik, Xiaodong Wen Background and purpose: Chronic hepatitis C (CHC) triggers oxidative stress, which is closely associated with emergence of hepatocellular carcinoma (HCC). On the other hand, hyperme-thylation-induced transcriptional inactivation of tumor suppressor genes (TSGs) has been reported in HCC. The purpose of this study is to clarify the association between oxidative stress, epigenetic alterations and development of HCC in CHC patients.

The HCV/CyA/MMF

combination reduced cell viability by 2.8-fold, and increased clCas3 and clPARP by 18.3- and 32.7-fold respectively. A similar effect was seen with the combination of HCV/Tac/MMF and HCV/Sir/MMF. In HCV-infected PMoH exposed to either CyA, Tac or Sir, caspase inhibition with Q-VD improved cell viability buy Barasertib by 58%, 53%, and 43%, reduced clCas3 by 88%, 89%, and 83%, and reduced clPARP by 93%, 92%, and 92% respectively. These significant improvements with Q-VD were also seen in HCV-infected cells exposed to CyA or Tac or Sir + MMF. Conclusion: In HCV-infected hepatocytes, CyA, Tac and Sir were all found to increase hepatocyte apoptosis. These findings help explain the accelerated progression of liver disease in post-transplant HCV recurrence. The finding that Q-VD reversed hepatocyte cell death in this setting provides

a rationale for targeting apoptosis to reduce liver injury here. F HACZEYNI,1 A MRIDHA,1 M YEH,2 N TEOH,1 G FARRELL1 1Liver Research Group, ANU Medical School, The Canberra Hospital, ACT. 2Department of Pathology, University of Washington, Seattle, WA, USA Introduction/Aims: In unhealthy obesity, adipose inflammation is associated with Venetoclax nmr insulin resistance and non-alcoholic steatohepatitis (NASH). However, the mechanism of adipose inflammatory recruitment is unknown. We studied whether Toll-like receptor 9 (Tlr9) and intracellular adapter protein Myeloid differentiation factor (MyD) 88 (signalling intermediate of multiple TLRs) play a role in adipose inflammation and development of NASH. Methods: Female Tlr9-/-, MyD88-/-, and wildtype (WT) mice (n = 6 − 14) on C57BL/6J (B6) strain were fed either rodent chow or atherogenic

(Ath) (SF03-020; Glen Forrest) diet from weaning to 24 weeks of age. Fasting blood, liver, and periovarian white adipose tissue (Pov WAT) and other fat pads were removed under anaesthesia for tissue and molecular analyses. Results: Ath diet increased body weight in all groups of mice, but deletion of Tlr9 reduced weight gain (32 g ± 0.9 Tlr9-/- vs. 43 g ± 1.8 WT p < 0.05). Similarly, Pov WAT weight expansion was less in Ath-fed Tlr9-/- than corresponding WT and MyD88-/- mice. Conversely, MCE liver weights increased in Ath-fed WT mice but significantly less in both Ath-fed Tlr9-/- and MyD88-/- counterparts. While grade 2–3 steatosis occurred in all Ath-fed mice, necroinflammatory score (Fig A) and NAFLD activity score (NAS) were less in Tlr9 and MyD88 deleted mice. Surprisingly, serum ALT levels were higher in Tlr9-/- and MyD88-/- than WT mice, even in those fed chow (p < 0.05) (Fig B). Further, ser insulin was higher in Ath-fed MyD88-/- mice than other groups, albeit fasting blood glucose levels were similar in all groups. In Pov WAT, adiponectin mRNA levels decreased with Ath dietary feeding in all groups compared to chow.

Furthermore the topical steroid budesonide is now being evaluated as an alternative to prednisone or prednisolone in order to achieve or maintain remission with less steroid specific side effects.366-369 Retrospective analyses have indicated that the long-term maintenance therapies need not be life-long.347 Twelve percent of patients treated with these schedules are able to be permanently CYC202 nmr withdrawn from medication after 69 ± 8 months of follow-up, and the probability of a sustained remission after total drug withdrawal is 13% after 5 years.347 These observations justify periodic attempts at drug withdrawal in all patients with longstanding (≥12 months) inactive disease.

The inability to discontinue azathioprine mandates indefinite treatment. Relapse in children is characterized by any manifestation of recrudescent hepatic inflammation after drug withdrawal.35,36,279-281,283,305,358-361 Its

frequency in children is the same or higher than that observed Navitoclax ic50 in adults. Relapse is often associated with nonadherence to treatment.370 The occurrence of relapse in children justifies reinstitution of the original treatment regimen. Indefinite low-dose therapy can then be instituted after suppression of disease activity using prednisone in combination with azathioprine or 6-mercaptopurine. Maintenance therapy with azathioprine alone is a management option for children who have relapsed.305 Recommendations: 31. The first relapse after drug withdrawal should be retreated with a combination of prednisone plus azathioprine at the same treatment regimen as with the initial course of therapy and then tapered to monotherapy with either azathioprine (2 mg/kg daily) as a long-term maintenance therapy or with indefinite low dose prednisone (≤10 mg daily) in patients intolerant

of azathioprine. (Class IIa, Level C) 32. Gradual medchemexpress withdrawal from long-term azathioprine or low-dose prednisone maintenance therapy should be attempted after at least 24 months of treatment and continued normal serum AST or ALT level only after careful benefit risk evaluation in patients who had previously relapsed. (Class IIa, Level C) Treatment failure should be managed with high dose prednisone (60 mg daily) or prednisone (30 mg daily) in combination with azathioprine (150 mg daily) before considering other drugs such as cyclosporine, tacrolimus, or mycophenolate mofetil. Alternative medications that have been used empirically for treatment failure in adults have included cyclosporine,308,371-376 tacrolimus,377-379 ursodeoxycholic acid,380 budesonide,381 6-mercaptopurine,382 methotrexate,383 cyclophosphamide,384 and mycophenolate mofetil.357,385-391 In each instance, experiences have been small and anecdotal. Only ursodeoxycholic acid has been evaluated by randomized controlled clinical trial,380 and it and budesonide are the only salvage therapies in which the reported experiences have been negative.

Informed consent was obtained from all patients. Data on life-long treatment history, 5-year data on bleeds, clotting

factor consumption and socioeconomic parameters were assessed from patient files and treatment records. Assessment of outcome included physical examination by a physiotherapist using the HJHS (Haemophilia Joint Health Score 1.0, max 148 points) [22] and questionnaires to assess self-reported activities (HAL, max 100 points)[23], physical activity expressed in METS (IPAQ) [24] and quality of life by the SF36 and the Euroqol (EQ-5D)[25], to allow the calculation of costs and utilities. To establish validity of joint evaluation by the HJHS, a Epigenetic Reader Domain inhibitor training session was held including physiotherapists from each centre and 12 patients eligible for the study. The Intra Class Correlation between physiotherapists was good at 0.80. Treatment, clinical outcome, clotting factor consumption and socioeconomic parameters compared between strategies will be analysed. These Alectinib results are expected to provide more insight into the long-term consequences of these different prophylactic treatment strategies. (Dr Miners) Severe haemophilia is a lifelong

disease requiring treatment with exogenous clotting factor. Where available, treatment is typically either given on demand, following a bleed, or prophylactically with the aim of preventing bleeding in the first instance. Prophylaxis is usually defined as being ‘primary’, initiated medchemexpress before the onset of serial bleeding, or ‘secondary’, when treatment is started sometime after this process has begun. Prophylaxis is considered to be the clinical treatment of choice but it is costly to provide. However, the provision of costly treatments is justified on economic grounds if it also generates proportional increases in (health) benefits. In non-market situations, typically such as the provision of health care, information on cost effectiveness is generated by performing economic evaluations, where the costs and

benefits of two or more health technologies are compared. The results from economic evaluations help decision makers to allocate resources towards health care technologies that are considered to be cost effective, or efficient, and away from those that are not. Although a number of economic evaluations of prophylaxis have been published, they report a broad array of results ranging from being ‘dominant’ (where prophylaxis is considered less costly and more beneficial than treatment on demand) to prophylaxis costing over €1 million per additional quality-adjusted life-year (QALY). Thus, there is a considerable amount of uncertainty in the evidence base. Given that all (health care) resources are finite, and demand will always outstrip supply, the production and use of information on cost effectiveness will, or at least should, always be an important component of decision-making.

From a survey of Fusarium species associated with maize ear rot in nineteen provinces in 2009 in China, ten strains isolated from Guizhou and Hubei provinces were identified as F. temperatum. Morphological and molecular phylogenetic analyses based on the DNA sequences of individual translation elongation factor 1-alpha and β-tubulin genes revealed that the recovered isolates produced

macroconidia typical of four-septate with a foot-shaped basal cell and belonged to F. temperatum that is distinctly different from its most closely related species F. subglutinans Pifithrin �� and others within Gibberella fujikuroi complex species from maize. All the strains from this newly isolated species were able to infect maize and wheat in field, with higher pathogenicity on maize. Mycotoxin

determination of maize grains infected by the strains under natural field condition by ultra-high-performance liquid chromatography–tandem mass spectrometry and gas chromatography–mass spectrometry analyses showed that among fifteen mycotoxins assayed, two mycotoxins fumonisin B1 and B2 ranging from 9.26 to 166.89 μg/g were detected, with massively more FB2 mycotoxin this website (2.8- to 108.8-fold) than FB1. This mycotoxin production profile is different from that of the Belgian population in which only fumonisin B1 was barely detected in one of eleven strains assayed. Comparative analyses of the F. temperatum and F. subglutinans strains

showed that the highest fumonisin 上海皓元 producers were present among the F. temperatum population, which were also the most pathogenic to maize. These results suggested a need for proper monitoring and controlling this species in the relevant maize-growing regions. “
“A Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) assay was employed to develop a simple and efficient system for the detection of Zucchini yellow mosaic virus (ZYMV) in squash and melon plants. The RT-LAMP assay took 30 min under isothermal condition at 64°C by employing a set of four primers targeting ZYMV. The sensitivity of RT-LAMP was 10-fold greater than that of the RT-PCR assay in the detection of ZYMV in infected tissues of squash and melon. No reaction was detected from the tissues of healthy plants by either RT-LAMP or RT-PCR assay. The RT-LAMP product of the tested samples can be visualized by staining directly in the tube with SYBR Green I dye. The sensitivity of SYBR Green I staining method is similar to that analyzed by gel electrophoresis. Field-grown squash and melon plants were tested using RT-PCR and RT-LAMP. Both RT-LAMP and PCR could detect ZYMV in symptomatic or symptomless tissues of infected plants. However, the RT-LAMP assay is superior to RT-PCR because it is rapid, simple, and highly sensitive; therefore, RT-LAMP is a useful and practical method for detection of ZYMV in cucurbits.

(HEPATOLOGY 2012) Interleukin-33 (IL-33) or IL-1F11 is a recently described member of the IL-1 cytokine family which includes IL-1α, IL-1β, and IL-18.1 IL-33 is widely expressed in various tissues and the cellular sources of IL-33 are mostly endothelial and epithelial cells, as well as smooth muscle cells, keratinocytes, astrocytes, adipocytes, fibroblasts, monocytes, macrophages, hepatic and pancreatic stellate

cells, and hepatocytes.2-5 Once released from the cells, IL-33 mediates its cytokine functions by interacting with its specific heterodimeric receptor comprising ST2 (IL-1 receptor-like 1) and IL-1RAcP (IL-1 receptor accessory protein) in an CH5424802 research buy autocrine or paracrine manner.6, 7 IL-33 targets cells of the immune system mainly by way of the ST2 receptor. The ST2 is expressed by T helper 2 (Th2)-cells, basophils, eosinophils, natural killer (NK) cells, natural killer

T (NKT) cells, and dendritic cells. Functionally, IL-33 can act as a chemoattractant for Th2 lymphocytes and induce various proinflammatory cytokines or inflammatory mediators.5, 8 The expression of IL-33 has been clearly SCH 900776 chemical structure associated with many acute and chronic inflammatory diseases including arthritis, asthma, lung inflammation, allergy, Crohn’s disease, ulcerative colitis, sepsis, anaphylactic shock, and hepatitis.2, 3, 9 Recently, we and others have shown that the IL-33/ST2 axis could play an important MCE role in acute and chronic liver diseases.2, 3, 10 Surprisingly, we found that IL-33 is strongly expressed in hepatocytes and demonstrated that

NKT cells were responsible for regulating IL-33 in hepatocytes during concanavalin A (ConA)-induced acute hepatitis.2 Pretreatment of WT mice with recombinant IL-33 prevents the severity of ConA-induced liver damage by recruiting regulatory T-cells and CD4+ T-cells into the liver.10 However, the molecular mechanisms that trigger nuclear IL-33 expression in hepatocytes remain unknown. The administration of ConA in mice provides an experimental model of T-cell-mediated liver injury, which resembles viral or autoimmune hepatitis in humans.11, 12 In the ConA model, the cytotoxic effector molecules and their receptors like tumor necrosis factor alpha (TNFα), perforin-granzyme, FasL/Fas, and tumor necrosis factor related apoptosis inducing ligand (TRAIL)/DR5 play a crucial role for hepatitis development as well as hepatocyte cell death.13-25 Indeed, TRAIL and DR5 expression increase and liver injury is suppressed in TRAIL−/− mice or after blockage of the DR5 receptor, which suggests a critical role of TRAIL/DR5 in the pathophysiology of ConA-induced acute hepatitis.23, 24 In the present study, we aimed to better characterize the expression of IL-33 during acute hepatitis by using wildtype (WT), perforin−/−, TRAIL−/−, and CD1d−/− mice as well as in primary murine hepatocytes.

To quantify the percentage of cells with apoB-GFP-LC3 puncta, at least 200 cells per condition were counted in randomly selected fields. In all cases, only those cells with four or more prominent puncta of

apoB-GFP-LC3 were scored positively. At least three independent experiments were performed for each graph, unless otherwise indicated. The mean ± standard error of the mean is shown in figures. All statistical calculations were completed using GraphPad PRISM software (version 5). For grouped analyses, a two-way ANOVA was used followed by a Bonferroni post-hoc test. To compare control to different treatments a one-way ANOVA was applied followed by a Dunnett’s Multiple Comparison Test. Probability values of less than 0.05 were considered to be statistically significant. As a first approach to gain insight into the role of autophagy under ER stress conditions, we examined the colocalization of apoB with LC3 (the microtubule this website associated protein 1 light chain 3), an autophagosome marker. Colocalization of apoB with GFP-LC3 was barely detectable (Fig. 1A, panels a-c) under untreated conditions in McA-RH7777 cells transiently expressing GFP-conjugated LC3 (GFP-LC3) for 24 hours. However, the colocalization of apoB with GFP-LC3, referred to as apoB-GFP-LC3 puncta, was markedly enhanced following 4 mM GLS treatment for 4 hours (Fig. 1A, panel d-f). Increasing the GLS concentration

to 16 mM led to high levels of apoB-GFP-LC3 puncta find more concentrated in a juxtanuclear localization, and in the distal area near the plasma membrane (Fig. 1A, panels g-i). The density of apoB-GFP-LC3 puncta–positive cells as well as the number of apoB-GFP-LC3 puncta in each positive cell increased with rising concentrations of GLS (0-16 mM) (*P < 0.05) (Fig. 1B). Concomitantly, increased apoB-GFP-LC3 puncta MCE were correlated positively with the degradation of newly synthesized apoB in a GLS dose-dependent manner (*P < 0.05) (Fig. 1C). Moreover, as shown in Fig. 1E, under the basal (Fig. 1D, panel c), and TM-treated (Fig. 1D, panel f) or GLS-treated (Fig. 1D, panel i) conditions, the apoB-GFP-LC3 puncta–positive cells, and number of apoB-GFP-LC3 puncta was substantially increased by

a longer GFP-LC3 expression time (48 hours). We next sought to further investigate links between the induction of ER stress and the autophagic degradation of apoB. Experiments were performed in McA-RH7777 cells treated with TM (5 μg/mL) or GLS (5 mM) for 4 hours in the presence or absence of 4-phenyl butyric acid (PBA, 1 mM), a chemical inhibitor of ER stress.25 Treatment with TM or GLS resulted in increased apoB-GFP-LC3 puncta–positive cells and a higher number of apoB-GFP-LC3 puncta in each cell (Fig. 2A, panels f and i; and analysis of data shown in Fig. 2C; different letters indicate significance, P < 0.05). Similar results were obtained when colocalization of apoB and endogenous LC3 was examined in nontransfected cells (Fig. 2F, and Supporting Fig. 1).