Category: PLA

M. receptors (NLRs) and elicit an inflammatory response characterized by caspase-1 activation (40, 42). The NLR protein Nalp1b has also been linked to caspase-1 activation and macrophage cytolysis mediated by anthrax lethal toxin (LT) (6, 36). However, it is unclear how LT activates the proinflammatory protein Nalp1b and how this results in caspase-1 activation in murine macrophages. LT is considered the primary virulence factor produced by the gram-positive organism spores. LT is usually a protein toxin consisting of two subunits, protective antigen (PA) and lethal factor (LF) (10). PA binds to specific cell surface receptors and mediates endocytosis of LF, a zinc protease. The proteolytic activity of LF is essential for the cytopathic and lethal effects observed in LT-treated mice (15, 20). The response of murine macrophages to LT exposure is usually mouse strain dependent. Murine macrophages are either susceptible or resistant to LT-mediated caspase-1 activation and cytolysis (29, 30). Genetic mapping experiments have identified a single gene, alleles from susceptible murine strains in the resistant C57BL/6 background renders the resulting macrophages susceptible to rapid LT killing (6). Nalp1b belongs to the NLR family of intracellular surveillance proteins, which are able to recognize pathogen-associated molecular patterns, including lipopolysaccharide (LPS) (25, 34, 40). In contrast to murine Nalp1b, the human NLR proteins NALP1 and NALP3 have been well characterized (25, 40). Stimulation of NALP1 or NALP3 results in the recruitment of downstream components and the formation of the inflammasome complex, which appears to be a critical event associated with caspase-1 activation (1, 24, 26). The NOD of NALP proteins is required for dimerization, and the LRR domain name has a microbe-sensing function (40). The pyrin domain name (PYD) and the caspase recruitment domain name (CARD) of Rabbit polyclonal to GLUT1 NALP1 and NALP3 are essential for the recruitment of ASC and caspase-1, respectively (24, 26). In contrast to human NALP1, the PYD is usually absent in murine Nalp1b, and the involvement of murine Asc in Nalp1b inflammasome activation is usually therefore questionable (6). NLR stimulation by specific ligands results in activation of proinflammatory caspase-1 and cell death (11, 12, 16). Activated caspase-1 then processes pro-interleukin-1 (pro-IL-1), IL-18, and IL-33 into their mature forms (22, 37). Consistent with a role for Nalp1b in LT susceptibility, caspase-1 is usually activated in susceptible LT-treated macrophages but not in resistant cells (31). Studies with caspase-1-deficient murine macrophages LDS 751 and caspase-1 inhibitors suggest that caspase-1 is essential for LT killing of susceptible murine macrophages (6, 31, 41). The mechanism by which microbial components, including LT, activate the inflammasome and the way in which this results in caspase-1 activation are poorly comprehended. In contrast to bacteria, which LDS 751 contain multiple virulence factors that simultaneously activate several NLRs, LT is usually a single virulence factor and appears to represent an ideal LDS 751 LDS 751 model system to study microbial inflammasome induction and caspase-1 activation. Our findings indicate that LT triggers the formation of an inflammasome complex made up of Nalp1b and caspase-1 in murine macrophages. In untreated macrophages, caspase-1 was a part of low-molecular-weight fractions and shifted toward high-molecular-weight fractions following LT treatment. Formation of the high-molecular-weight complex, presumably the inflammasome, coincides with caspase-1 activation and macrophage lysis. Caspase-1-associated proteins also included caspase-11 and the caspase-1 target -enolase in LT-treated macrophages. MATERIALS AND METHODS Cell lines and plasmids. The BALB/c-derived murine macrophage cell line J774A.1 and the human kidney fibroblast line 293T were obtained from the American Type Tissue Culture Collection (ATCC, Manassas, VA). Cells were cultured in Dulbecco’s altered Eagle’s medium (DMEM) (Mediatech, Herndon, VA) made up of 10% fetal bovine serum (HyClone, Logan, UT) supplemented with 100 U/ml penicillin and 100 g/ml streptomycin (Gibco, Grand Island, NY). The procaspase-1 expression vector was purchased from ATCC. The.

We did not expect to see extensive co-localization of Dsg2 and Cav-1 since desmogleins are predominantly found in desmosomes and Cav-1 in lipid rafts. S3. Cav-1competing peptides induce loss of keratinocyte adhesion. A431 cells were incubated with 5 M AP or AP-Cav-1 in serum-free medium for 2 hr. Cell sheets were subjected to dispase-based keratinocyte dissociation assay showing more fragmentation after treatment with AP-Cav-1 peptides, as compared to control untreated or treated with AP alone. Shown are representative results from three separate experiments. NIHMS310516-supplement-3.jpg (78K) GUID:?F42984B7-CFEE-4C59-80FB-F10671D839C2 Abstract Desmoglein 2 (Dsg2) is a desmosomal cadherin that is aberrantly expressed in human skin carcinomas. In addition to its well-known role in mediating intercellular desmosomal adhesion, Dsg2 regulates mitogenic signaling that may promote cancer development and progression. However, the mechanisms by which Dsg2 activates these signaling pathways and the relative contribution of its signaling and adhesion functions in tumor progression are poorly understood. In this study we show that Dsg2 associates with caveolin-1 (Cav-1), the major protein of specialized membrane microdomains called caveolae, which functions in both membrane protein turnover and intracellular signaling. Sequence analysis revealed that Dsg2 contains a putative Cav-1 binding motif. A permeable competing peptide resembling the Cav-1 scaffolding domain bound to Dsg2, disrupted normal Dsg2 staining and interfered with the integrity of epithelial sheets in skin tumors from transgenic mice overexpressing Dsg2. Collectively, these data are consistent with the possibility that accumulation of truncated Dsg2 protein interferes with desmosome assembly and/or maintenance to disrupt cell-cell adhesion. Furthermore, the association of Dsg2 with Cav-1 may provide a mechanism for regulating mitogenic signaling and modulating DZNep the cell surface presentation of an important adhesion molecule, both of which could contribute to malignant transformation and tumor progression. null mice revealed that Dsg2 contributes to embryonic stem cell proliferation, particularly in the inner cell mass of the developing blastocyst (Eshkind et al., 2002). Dsg2 is aberrantly expressed in select epithelial malignancies, including squamous cell carcinomas (Biedermann et al., 2005; Harada et al., 1996; Kurzen et al., 2003). Similarly, genetic profiling of prostate cancer cell lines showed increased expression of Dsg2 in a metastatic cell line, as compared to its non-metastatic DZNep syngeneic precursor cell (Trojan et al., 2005). Dsg2 expression is also upregulated in squamous cell carcinoma (SCC) cell lines in comparison to cultured keratinocytes (Denning et al., 1998; Harada et al., 1996; Sch?fer et al., 1994). We recently showed that Dsg2 is highly expressed in malignant skin carcinomas, including squamous cell carcinomas, basal cell carcinomas, sweat and sebaceous gland carcinomas and adenocarcinomas (Brennan and Mahoney, 2009). Collectively, these results support a role for Dsg2 in epithelial cell growth, survival and malignant transformation. However, the mechanisms by which Dsg2 activates these signaling pathways and promotes tumor formation are unknown. Caveolins are a family of hairpin-like, DZNep palmitoylated, integral membrane proteins that oligomerize and bind to cholesterols and sphingolipids to form specialized areas of the membrane distinct from the clathrin-coated pits. The caveolins form flask-shaped invaginations of 50-100 nm in diameter called caveolae (Severs, 1988). There are three caveolin isoforms: Cav-1 ( and ), Cav-2 and Cav-3. While Cav-1 and Cav-2 are ubiquitously expressed, Cav-3 expression is predominantly restricted to muscle cells (Scherer et al., 1995; Tang et al., 1996). Caveolins and caveolae have been implicated as regulators of key cellular functions, including cholesterol transport and homeostasis (Fielding and Fielding, 1995; Smart et al., 1996), endocytosis and endocytic vesicle trafficking (Schnitzer and Oh, 1996), cell adhesion and apoptosis (Kurzchalia and Parton, 1999; Lisanti et al., 1994; Okamoto, 1998; Okamoto et al., 1998; Shaul and Anderson, 1998). Specific cell signals can be also transmitted through a spatially controlled organization of cell receptors into the caveolae. Indeed, the epidermal growth factor (EGF) receptor has been shown to stimulate the phosphorylation of Cav-1, thus enhancing caveolae assembly (Orlichenko et al., 2006; Severs, 1988; Simons and Toomre, 2000; Singer and Nicolson, 1972). Furthermore, Cav-1 is essential for integrin-mediated activation of PI3-K/AKT (Sedding et al., 2005). Conversely, overexpression of Cav-1 abrogates anchorage-independent cell survival (Engelman et al., 1997), and TSPAN2 suppresses cell growth (Lee et al., 1998). Additionally, Cav-1 binds to and inhibits kinases involved in mitogenic signaling pathways. Cav-1 expression can modulate Wnt/-catenin/Lef-1 signaling by regulating the intracellular localization of -catenin (Galbiati et al., 2000). Consistent with these findings, mounting evidence suggests that diseases associated with deregulated signaling pathways often result from aberrant expression or localization of Cav-1. In cancer, the role for Cav-1 is complex, as it serves both as a modulator of tumor suppression as well as oncogenesis. Mutations in the gene have been linked to human breast cancer, suggesting that loss of Cav-1 function plays a significant role in tumor initiation (Chen et al.,.

After removal of the cumulus cells, the denuded oocytes were placed back in their original wells containing standard maturation medium without or with 250?M stearic acid. were exposed to saturated stearic acid during maturation. SCD inhibition in the presence of stearic acid significantly reduced the developmental competence of oocytes and increased the incidence of apoptosis in cumulus cells. The esterified oleic/stearic acid ratio of the neutral lipid fraction in cumulus cells decreased in the presence of SCD inhibitors when COCs were exposed to saturated free fatty acids during maturation, indicating the SCD-specific conversion of saturated fatty acids under noninhibiting conditions. The observation that Rabacfosadine cumulus cells can desaturate the Rabacfosadine potentially toxic stearic acid into oleic acid via SCD activity provides a mechanistic insight into how the cumulus cells protect the oocyte against toxicity by saturated fatty acid. [22, 23], while the human and bovine genomes only contain two SCD genes: and [24, 25]. The aim of the current study is to determine how cumulus cells protect the oocyte against free fatty acids. Materials and methods Chemicals Unless stated otherwise, all chemicals used were obtained from Sigma Chemical Co (St. Louis, MO, USA) and were of the highest purity available. Solvents (acetone, acetonitrile, chloroform, methanol, and hexane) were of high-performance liquid chromatography (HPLC) grade (Labscan, Dublin, Ireland). Collection and maturation of cumulus-oocyte complexes Bovine ovaries were collected at a slaughterhouse and transported to the laboratory within 2 h of slaughter. Approval of an independent ethical committee was not needed as ovaries were a rest product of the regular slaughter process in the slaughterhouse. Antral follicles between 2 and 8 mm in diameter were aspirated by means of a suction pump under low vacuum. The follicular aspirates were pooled in a conical tube and allowed to settle for 15 min. Oocytes with a multilayered cumulus investment were selected from the follicular fluid, washed three times in HEPES-buffered M199 (Gibco BRL, Paisley, UK), and randomly allocated in groups of 35C70 COCs per well in four-well culture plates (Nunc A/S, Roskilde, Denmark). In vitro Rabacfosadine maturation (IVM) was carried out for 23 h according to our standard protocol [5] in maturation medium consisting of 500?l M199 per well, and no coverage of oil, supplemented with 26.2 mM NaHCO3, 0.02 IU/ml FSH (Sioux Biochemical Inc., Sioux Center IA, USA), 0.02IU/ml LH (Sioux Biochemical Inc.), 7.7?g/ml cysteamine, 10?ng/ml epidermal growth factor, and Rabbit polyclonal to ADAM18 1% (v/v) penicillin-streptomycin (Gibco BRL) at 39C in a humidified atmosphere of 5% CO2 in air. Fertilization and embryo culture After 23 h of maturation, in vitro fertilization was performed with 0.5 106/ml sperm from a bull with proven fertility. At 18C22 h after sperm addition, the cumulus cells and adhering sperm cells were removed by vortexing the presumed zygotes for 3 min in the experiment with SCD inhibition. Subsequently, the zygotes were transferred in groups of 35C70 to wells with 500?l pre-equilibrated synthetic oviductal fluid (SOF, [26]). Fertilization and embryo culture were performed, according to our standard procedure [5], in a humidified incubator at 38.5C with 5% CO2 and respectively 20% and 7%O2. At day 5 of culture, cleaved embryos were transferred to fresh SOF and cultured until day 8 on which embryonic development was assessed. In vitro maturation with free fatty acids and stearoyl-CoA desaturase 1 inhibitor The free fatty acids used in the maturation experiments were processed according to our standard protocol [5] and bound to 100% delipidified bovine serum albumin resulting custom-tailored lipidified BSA, and were stored in stock at a concentration of 10 mM bound to 10% (w/v) fatty acid-free BSA (fatty acid:BSA stoichiometry of 5:1). For the experiments assessing the importance of cumulus cells in protecting oocytes against free fatty acids, the cumulus cells of oocytes were removed after a maturation period of 8 Rabacfosadine h by vortexing COCs for 3 min in HEPES-buffered M199 [27]. After removal of the cumulus cells, the denuded oocytes were placed back in their original wells containing standard maturation medium without or with 250?M stearic acid. As a control, groups of intact COCs were matured in maturation medium without or with 250?M stearic acid. For the experiment where cumulus cells were removed at 8 h, cumulus cells from the control group with maturation as intact COCs were removed before fertilization by vortexing for 3 min, which is a modification from our standard procedure. Oocytes were fertilized and cultured according.

Supplementary MaterialsFigure S1: HT1080 cells or HT1080 overexpressing hMR1 (HT1080-hMR1) cells were cultured in the current presence of increase multiplicity of infection (MOI) of Escherichia coli (Ec). quantities and HLA-DR manifestation. The two populations (V7.2+CD161? and V7.2?CD161+ cells) showed no differences in percentage or activation markers comparing base line (BL) and days after vaccine ingestion (D7, D9 and D11) or between Placebo, Non-Responders and Responders.(PDF) ppat.1003681.s003.pdf (95K) GUID:?0604EDBA-424B-43DA-883C-A74AC332C1E0 Movie S1: MAIT cells were seeded about Hela cells and imaged every 3 min for 15 hours.(MOV) ppat.1003681.s004.mov (24M) GUID:?9939018C-6BBF-40B1-912D-8B8305BC6469 Movie S2: MAIT cells were seeded on hMR1 overexpressing Hela cells in the presence of bacteria lysate and imaged every 3 min for 15 hours.(MOV) ppat.1003681.s005.mov (16M) GUID:?360A2351-D003-4ED7-8B27-1DE63D31F87F Movie S3: MAIT cells were seeded about hMR1 overexpressing of Hela cells and imaged every 3 min for 15 hours.(MOV) ppat.1003681.s006.mov (16M) GUID:?13A21325-CECD-48B0-9C94-0A1410C077EC Movie S4: MAIT cells were seeded about hMR1 overexpressing Hela cells in the KBTBD7 presence of bacteria lysate and imaged every 3 min for 15 hours. Rounding of the much bigger epithelial cells indicating death is frequent only when both hMR1 and the bacteria lysate are present.(MOV) ppat.1003681.s007.mov (13M) GUID:?348262DC-F393-4455-874B-83DAA4E60530 Abstract Mucosal associated invariant T cells (MAIT) are innate T lymphocytes that detect a large variety of bacteria and yeasts. This acknowledgement depends on the detection of microbial compounds presented from the evolutionarily conserved major-histocompatibility-complex (MHC) class I molecule, MR1. Here we display that MAIT cells display cytotoxic activity towards MR1 overexpressing non-hematopoietic cells cocultured with bacteria. The NK receptor, CD161, highly indicated by MAIT cells, modulated the cytokine but not the cytotoxic response induced by bacteria infected cells. MAIT cells will also be triggered by and destroy epithelial cells expressing endogenous levels of MRI after illness with the invasive bacteria In contrast, MAIT cells were not triggered by epithelial cells infected by in a process requiring endogenous MR1, while the closely related bacterium is not. Upon acknowledgement, infected epithelial cells are efficiently lysed by MAIT cells. We also display the triggering of CD161, a natural killer receptor highly indicated by MAIT cells, can modulate the cytokine but not the cytotoxic function of these cells. Finally, we provide evidence that MAIT cells are triggered during the course of an experimental enteric Fedovapagon illness in humans. Our study provides important insight within the antibacterial function of MAIT cells and their connection with pathogenic bacterial varieties. Introduction Detection of bacterial infections occurs through detection of microbial compounds from the innate immune system receptors [1], [2]. As chlamydia advances, the adaptive disease fighting capability respond to substances made by these pathogens in an activity that will require Fedovapagon priming of na?ve cells and following differentiation and proliferation. Innate like T cells Fedovapagon bridge both of these systems by giving immediate effectors features in response towards the an infection [3], [4]. Certainly, as opposed to typical T cells that exhibit a very different T cell receptor (TCR) repertoire and so are limited by polymorphic MHC substances, innate like T cells screen semi-invariant TCRs and so are limited by non-polymorphic MHC-Ib substances. In human beings, they represent huge oligoclonal expansions with instant effector properties. Inside the innate-like T cells, Mucosa-Associated Invariant T (MAIT) cells are limited with the evolutionarily conserved MHC related molecule, MR1 [5], [6]. In human beings, MAIT cells are loaded in peripheral liver organ and bloodstream, are uniformly storage and screen a tissue-targeted phenotype [7], [8]. MAIT cells exhibit transcription elements connected with particular effector actions such as for example ZBTB16 and RORt [7], [8]. Appropriately, they exhibit at their cell surface area high degrees of cytokine receptors for IL-18, IL-23 and IL-12.

Supplementary Materials1. particular for epitopes of HCMV phosphoprotein-65, tetanus toxoid precursor, Epstein-Barr pathogen nuclear antigen 2 or HIV gag proteins. Inflated DYS-specific Compact disc4+ T cells comprised effector memory space or effector memory-RA+ subsets with limited TCR-beta utilization and almost monoclonal CDR3 including novel conserved proteins. Expression of the near monoclonal TCR inside a Jurkat cell transfection program validated good DYS specificity. Inflated cells had been polyfunctional, not really senescent, and shown high degrees of granzyme-B, CX3CR1, Compact disc38 or HLA-DR, but were less Compact disc38+HLA-DR+ co-expressing frequently. The inflation system didn’t involve apoptosis suppression, improved proliferation or HIV gag cross-reactivity. Instead, the findings suggest that intermittent or chronic expression of epitopes such as DYS drive inflation of activated CD4+ T cells that home to endothelial cells and have the potential to mediate cytotoxicity and vascular disease. Introduction Classical CD4+ and CD8+ memory T cell responses against viruses expand during primary 3-Aminobenzamide infection and contract to low 3-Aminobenzamide magnitudes after contamination resolution (1). However, CD8+ T cell responses to select epitopes of human (HCMV) (2, 3), rhesus (4), and murine cytomegalovirus (MCMV) (5C9) persist for decades at very high magnitudes after primary contamination or during latency. This phenomenon is usually termed memory inflation and has been best characterized among CMV-specific CD8+ T cells that consist of mainly CD45RO+ CCR7? CD27? T cells (effector memory/TEM) and their CD45RA+ revertants, CD45RO? CCR7? CD27? T cells (effector memory-RA+/TEMRA) (8C12). CMV-specific CD8+ T cells express high levels of CX3CR1 that bind CX3CL1 (fractalkine), which is usually expressed on vascular endothelial cells (VECs), a major target of CMV latent contamination (1). Classical CMV-specific CD8+ T cells display an IL-7-receptor-alpha/CD127+ programmed cell death protein-1?, PD-1? phenotype (capable of homeostatic proliferation controlled by IL-7 and other cytokines), while inflated CMV-specific CD8+ T cells are CD127? PD-1? T cell immunoglobulin and ITIM domain name/TIGIT? Granzyme B+ CX3CR1+ with evidence suggesting they are maintained by low-level exposure to persistent antigen from stochastic CMV reactivation (1, 13C16). These data suggest inflated responses are maintained through recurrent stimulation by peptide-MHC (17C19) produced by persistent, stochastic expression of specific CMV transcripts (20C22). These epitopes are presented to CMV-specific T cells by latent HCMV-infected, non-hematopoietic reservoirs, including VECs, lymph node (LN) stroma cells, and cells in the bone marrow and lungs (1, 23C25). Maintenance of inflated CMV-specific T cell responses might also depend on their longer telomeres that positively correlate with persistence (26), or on epitope cleavage by constitutive proteasomes (6, 27). CMV-specific CD4+ T cells suppress HCMV Rabbit polyclonal to SP3 lytic replication (28) and maintain CD8+ T cell inflation (29). HCMV lysate-specific CD4+ T cells persist at high magnitudes in HIV+ 3-Aminobenzamide HCMV+ co-infection (30), which might be due to higher HCMV disease burden (31, 32). Yet it is not known whether CD4+ T cells specific to individual HCMV epitopes undergo memory inflation in co-infected subjects. Glycoprotein B/gB has the highest population prevalence of CD4 responses of any HCMV protein (33). gB polyprotein colocalizes to endosomes that process and present its class II epitopes directly from infected endothelial cells upon IFN–induced HLA class II expression (28, 34, 35) without needing professional APCs. gB-loaded endosomes are also secreted as immunogenic exosomes that stimulate CD4+ memory T cells (36, 37). In HLA-DRB1*07:01 (DR7+) persons, the most immunogenic gB epitope is the extremely conserved DYSNTHSTRYV (DYS) epitope that is recognized by cytotoxic, CX3CR1+ CD4+ T cells (11, 38). HIV+ HCMV+ co-infection is usually implicated in the emerging higher incidence of HCMV-related, non-AIDS comorbidities of cardiovascular diseases including hypertension, coronary 3-Aminobenzamide artery disease, and stroke despite suppressive antiretroviral therapy (ART) (31, 39C43). These disease risks are further increased in co-infected subjects with elevated CD4+ T cell 3-Aminobenzamide activation (CD38+HLA-DR+) (44), which are mostly CMV-reactive (45) and are reduced by anti-CMV.

Glycosyltransferases represent a large family of enzymes that catalyse the biosynthesis of oligosaccharides, polysaccharides, and glycoconjugates. the ganglioside biosynthesis pathway (Fig. 1). It really is proposed that reduced ST3GAL2 and B3GALT4 manifestation potential clients to vulnerability of dopaminergic neurons via aberrant ganglioside synthesis. In keeping with this hypothesis, the Bupranolol amount of GM1 ganglioside-expressing cells in the Bupranolol Parkinsons disease substantia nigra Bupranolol are decreased (Wu and (Fig. 1). In keeping with impaired ganglioside synthesis, the same research reported decreased ganglioside concentrations within both diseased human being caudate as well as the mouse striatum. It ought to be mentioned that despite significant homology to can be traditionally connected with are connected with familial ALS (Cooper-Knock pursuing Bupranolol in an impartial genome-wide display for DNA mutations in 12 trios including sporadic ALS individuals and unaffected parents (Pamphlett = 0.007). Identical testing didn’t determine an enrichment of ALS-associated mutations within OGT, certainly we only determined two rare missense mutations within OGT in 4493 sporadic ALS patients. It should be noted that OGT is encoded on the X chromosome and therefore males are necessarily hemizygous, which may predispose to a neurodevelopmental phenotype rather than a late age-of-onset disease: for example mutations within N-terminal tetratricopeptide repeats of OGT are associated with X-linked intellectual disability (Gundogdu = 0.91). Table 2 Mutations in EOGT found in ALS patients that is interesting, but rather upstream changes in glycosyltransferase function that initiate toxicity. With this in mind we have highlighted genetic associations between mutations in glycosyltransferases and neurodegenerative disease. Most prominently we have recently discovered autosomal dominant mutations in GLT8D1 to be a monogenic cause of ALS. Disease-associated mutations have also been discovered in UGT8 and ST6GAL1; and we have revealed a new association between ALS and mutations in EOGT. Glycosyltransferases are likely to be an important therapeutic target in the effort of develop disease-modifying therapies for neurodegenerative disease. Acknowledgements The authors would like to thank the Project MinE GWAS Consortium. We are very grateful to those ALS patients and control subjects who donated biosamples. Funding We acknowledge grants from the Academy of Medical Sciences, EU Framework 7 Bupranolol (Euro-Motor), and the JPND/MRC SOPHIA, STRENGTH and ALS-CarE projects. T.M. is supported by the University of Sheffield Lee Newton PhD studentship. J.C.-K. holds a NIHR Clinical Lectureship and P.J.S. is supported as an NIHR Senior Investigator. This work was also supported by the NIHR Sheffield Biomedical Research Centre for Translational Neuroscience and the Sheffield NIHR Clinical Research Facility. Competing interests The authors report no competing KLF4 interests. Glossary AbbreviationsALS = amyotrophic lateral sclerosisO-GlcNAc = O-linked –N-acetylglucosamine.