Project description:Lignocellulosic biomass is composed of three major biopolymers: cellulose, hemicellulose and lignin. Although lignin has long been considered a waste product in biomass conversion efforts, its utilization has since been identified as critical to the economic viability of second-generation biofuel production. There is thus increasing interest in finding enzymes and enzyme cocktails which can efficiently deconstruct both the cellulose/hemicellulose and lignin components of lignocellulosic biomass. Analytical tools capable of quickly detecting both glycan and lignin deconstruction could are needed to support the development and characterization of efficient enzymes/enzyme cocktails.
Project description:The total protein of bone marrw myeloma cells was extracted with cell lysis buffer for IP. YBX1 antibody was used to perform Immunoprecipitation.
Project description:We investigated the molecular pathogenesis of LBW rats obtained by intraperitoneal injection of dexamethasone into pregnant animals. Normal-birth-weight (NBW) rats were used as controls with seven animals per group. When the rats were four weeks old, the left kidneys were removed and used for comprehensive label-free proteomic studies
Project description:URI keeps low levels of p53 in a TRIM28-MDM2 dependent manner, maintains SCD1 activity and accumulation of MUFAs, and subsequently promotes resistance to TKIs in cancer cell. URI-p53-SCD1 axis mediates resistance of TKIs and may explain why p53-wild type HCC still showed intrinsic resistance to TKIs. Moreover, the combination therapy identified here may represent a promising strategy for the approximately 41% of patients with advanced HCC who have wild-type p53 and high levels of URI/SCD1.
Project description:Human embryonic stem cells with one allele of the NEUROG3 gene genetically modified into a fusion gene NEUROG3-LINKER-TAGRFPT-P2A-EGFP-NLS were differentiated into the pancreatic lineage until Stage 4 Day 1 of the in vitro differentiation protocol (Rezania et al., 2014, as modified in Petersen et al., 2017). The cells were index-sorted according to their GFP expression to enrich for low GFP and high GFP cells, and deep single-cell RNA-seq was performed using the plate-based Smart-seq2 platform (Picelli et al. 2014).
Project description:The "prion-like" features of Alzheimer’s disease (AD) tauopathy and its relationship with amyloid β (Aβ) has never been experimentally studied in primates phylogenetically close to humans. We injected 17 macaques in the entorhinal cortex with nanograms of tau proteopathic seeds purified from AD brains or control extracts from aged-matched healthy brains, with or without intracerebroventricular co-injections of recombinant Aβ oligomers. After neuropathological examination of the macaque’s brain, we also performed mass spectrometry-based proteomics of macaques’ entorhinal cortex and CA1 to study the spatial response (local and distant) to tau seeds injections (and its modulation by oligomeric Aβ).
Project description:Extracellular vesicles (EV) as drug delivery nanocarriers are under intense investigation. Although clinical-grade EVs have been produced on a large-scale, low yield and high production costs of natural EVs (nEV) limit the relevant industrial translation. Recent studies show that mechanical extrusion of cells can generate nEV-like engineered EVs (eEV) which can also be used as drug nanocarriers. Moreover, in comparison with nEVs, eEVs have similar physicochemical properties. Nevertheless, a comprehensive comparison of cargo between nEVs and eEVs has not been investigated yet. Therefore, the aim of this study is to profile and compare eEVs to nEVs. Our results show that no significant difference was found in size, morphology, and classical markers between nEVs and eEVs derived from MDA-MB-231 cells. Protein sequencing data reveals the similarity of membrane proteins between the two groups was ~71%, while it was ~21% when pertaining to total protein cargo. Notably, a high similarity of membrane proteins was also found between nEVs and eEVs derived from eight additional cancer cell lines. Moreover, analysis of the top 1000 small RNA with RNA sequencing showed a ~65% similarity between the two groups. Altogether, we infer from the high similarity of membrane proteins and small RNA cargo that eEVs can be a good substitute for nEVs. In brief, our findings support previous studies which discovered eEVs own comparable performance of nEVs and could pave the way for clinical implementation of eEV-based therapeutics in the future.
Project description:The trans-Golgi Network (TGN) in plants is a main sorting hub for proteins and lipids. The lipid composition at the TGN is instrumental to the polar sorting of the auxin efflux carrier PIN2 from TGN to the apical membrane. Here, we immuno-purified the full and intact TGN using the TGN-resident syntaxin SYP61 to identify and quantify TGN-proteins which abundance at TGN is dependent on the acyl-chain length of very-long chain fatty acids-containing lipids such as sphingolipids.
Project description:Despite the progress in safety and efficacy of cell therapy with pluripotent stem cells (PSCs), the presence of residual undifferentiated stem cells or proliferating neural progenitor cells (NPCs) with rostral identity has remained a major challenge. Here we reported the generation of an LMX1A knock-in GFP reporter human embryonic stem cell (hESC) line that marks the early dopaminergic progenitors during neural differentiation. Purified GFP positive cells in vitro exhibited expression of mRNA and proteins that characterized and matched the midbrain dopaminergic identity. Further proteomic analysis of enriched LMX1A+ cells identified several membrane associated proteins including CNTN2, enabling prospective isolation of LMX1A+ progenitor cells. Transplantation of hPSC-derived purified CNTN2+ progenitors enhanced dopamine release from transplanted cells in the host brain and alleviated Parkinson’s disease symptoms in animal models. Our study establishes an efficient approach for purification of large numbers of hPSC-derived dopaminergic progenitors for therapeutic applications.