Tag: pbs

Strains and growth conditions All strains and plasmids used in this work are described in Supplementary Table 1. The clinical isolates were obtained from the Cystic Fibrosis Foundation Isolate Core at Seattle Children’s Hospital and were originally isolated from two different cystic fibrosis patients. Distribution of these isolates is covered by…

Ethics statement All animal studies were performed in accordance with the recommendations in the Canadian Council on Animal Care Guidelines. The reporting of this data in the manuscript follows the recommendations in the ARRIVE guidelines. The University of Calgary Health Sciences Animal Care Committee approved all animal protocols and surgical…

Amino acid sequence comparisons between GAS and GBS Cas9 identify orthologous active sites Relationships between the amino acid sequence of GBS Cas9 endonuclease and its molecular functions can be deduced from detailed studies of its GAS ortholog, SpyCas9 (Supplementary Fig. 1). As a first step in locating DNA complementary to a…

Ethics The Australian Genomics Acute Care study has Human Research Ethics Committee approval (HREC/16/MH/251). Parents provided informed consent for participation in the study, following genetic counseling. Study design and participants The Acute Care Genomics program is a national multi-site study delivering ultra-rapid genomic testing to critically ill pediatric patients with…

Wanyang Dong,1,* Qi Peng,1,* Xiaohua Tang,1,2,* Tian Zhong,1 Shunan Lin,1 Ziling Zhi,1 Jingyi Ye,1 Bixia Yang,1 Ning Sun,1,3 Wenchang Yuan1 1Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases, KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 510180, People’s Republic of China; 2Third Affiliated Hospital…

Expression and purification of human PTH1R The plasmid encoding human PTH1R (GenBank identifier: U17418.1; residues 27–491) was constructed and purified as previously reported17. The construct was expressed in HEK293 GnTI (N-acetylglucosaminyltransferase I-negative) cells (American Type Culture Collection, CRL-3022) using the BacMam system (Thermo Fisher Scientific), and the cells were grown…

NOTCH3 anti-peptide antibodies NOTCH3 is a transmembrane receptor (Fig. 1A) that plays multiple roles in development, homeostasis, and pathology. Mutations in NOTCH3 are responsible for the most common cause of inherited stroke and vascular dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL)8,9. Figure 1 Generation and characterization of…

Introduction The definition of an individual organism, traditionally defined by the invariable presence of a physiological unit and genetic homogeneity, is challenged by situations such as intraorganismal genetic heterogeneity (IGH), which refers to the presence of more than one genotype in a single organism (Schweinsberg et al., 2015). Cells within a…

Cell lines Human melanoma cell lines (A375 and A875) were kindly donated by the Department of Dermatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, 100 μg/ml penicillin and 100 μg/ml streptomycin at 37 °C in a…

Introduction Interleukin (IL)-2 was the first Food and Drug Administration approved immunotherapy for use in patients with metastatic melanoma and renal cell carcinoma.1 2 However, the use of recombinant IL-2 (Proleukin/aldesleukin) has been problematic due to low response rates, off-target effects, and severe dose-dependent toxicities. Due to its potent T-cell…

We present a protocol for assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) specifically on adipocytes using nucleus sorting with adipose tissues isolated from transgenic reporter mice with nuclear fluorescence labeling. ATAC-seq is a powerful technique to understand gene expression regulatory mechanisms. However, adipose tissue is difficult using this technique…

Abstract De novo protein design enhances our understanding of the principles that govern protein folding and interactions, and has the potential to revolutionize biotechnology through the engineering of novel protein functionalities. Despite recent progress in computational design strategies, de novo design of protein structures remains challenging, given the vast size…

Sample collection and cDNA synthesis In this study, we used porcine alveolar macrophages (PAMs) that were isolated from 16 different animals between the years 2011 and 2022, and subsequently stored. The PAMs were isolated by bronchoalveolar lavage (BAL) from lungs of euthanized 3-week-old healthy conventional piglets from a PRRSV negative…

NLRP3 expression and purification Both wild-type NLRP3 and CAPS mutants were cloned in the mammalian expression vector pcDNA3.1HisB. Plasmid was purified using Qiagen Giga Prep up to 2000 ng/μl and subsequently transfected into Expi293 with Expifectamine per manufacturer instructions. Expression was allowed to proceed for 3 days before harvesting. Cells were…

Cell lines, culture conditions, heat shock treatment and transfections HeLa, HEK293 and 2D12 (a NOA36-knockout HEK293-cell line generated in this work) cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 100 U/mL penicillin and 100 µg/mL streptomycin at 37 °C in a CO2 incubator. For heat…

Balance equation of any chemical reaction online for freeNO2 + H2O = HNO3 + NO =>3 NO2 + H2O = 2 HNO3 + NO (NH4)2CO3 = NH3 + H2O + CO2 =>(NH4)2CO3 = 2 NH3 + H2O + CO2 NaOH + H3PO4 = Na3PO4 + H2O =>3 NaOH + H3PO4…

Microarray-based expression analysis and GEO database analysis The DLBCL-related miRNA expression profiles of GSE29493 and GSE40239 were retrieved from the Gene Expression Omnibus (GEO) database (www.ncbi.nlm.nih.gov/geo/). MiR-124’s pan cancer analysis is conducted through online websites (www.picb.ac.cn/dbDEMC/). Bioinformatics predicted NFATc1 as the target of miR-124-3p The miRbase (www.mirbase.org/), miRDB (mirdb.org), and…

Study cohorts This study was conducted in compliance with the Declaration of Helsinki and was approved by the Maoming People’s Hospital (Approval No. 2021-hs-43) and Dongguan Ninth People’s Hospital (Approval No. 2022-8). COVID-19 was confirmed based on positive results of reverse transcription-polymerase chain reaction (RT-PCR) of nasopharyngeal samples. Whole blood…

Vector construction The Cas9/sgRNA vector (all-in-one) (GenBank accession no. OQ579018) contained sgRNA and Cas9-2A-mCherry expression cassettes. The sgRNA (5′-ATGCAGAACTAGAGTACAGC-3′) targets the phiC31 pseudo-attP intergenic site located on chromosome 3 of CHO-K1 genome (GenBank accession no. APMK01032147.1) (Fig. 1a). To construct the Cas9-mSA/sgRNA vector (GenBank accession no. OQ579019), the monomeric streptavidin (mSA)…

Product Specific Information Description: The human soluble IL-2 receptor alpha chain (sIL-2Ra) is generated by proteolytic cleavage of the cell-surface bound receptor on activated T cells. The IL-2Ra (also known as CD25) is expressed by early progenitors of the T and B lineage, as well as by activated mature T…

1. Introduction Tuberculosis (TB) is a significant global burden and is widely reported to be a major public health and economic problem, costing the world $617 billion between 2000 and 2015 and projected to cost $1 trillion between 2015 and 2030 (1). It is the second leading cause of death…

This protocol describes the fabrication and characterization of a photoresponsive prodrug-dye nanoassembly. The methodology for drug release from the nanoparticles by light-triggered disassembly, including the light irradiation setup, is explicitly described. The drugs released from the nanoparticles following light irradiation exhibited excellent anti-proliferation effects on human colorectal tumor cells. This…

In this study, a rat model of induction of periodontitis is presented via a combination of retentive ligature and repetitive injections of lipopolysaccharide derived from Porphyromonas gingivalis, over 14 days around the first maxillary molars. The ligation and LPS injection techniques were effective in inducing peridontitis, resulting in alveolar bone…

This protocol describes the isolation of mouse preadipocytes from subcutaneous fat, their differentiation into mature adipocytes, and the induction of insulin resistance. Insulin action is evaluated by the phosphorylation/activation of members of the insulin signaling pathway through western blot. This method allows direct determination of insulin resistance/sensitivity in primary adipocytes….

货号： CSB-PA008246 规格： 图片： 其他： 产品详情 Uniprot No.： 宿主： Rabbit 反应种属： Human,Mouse,Rat 免疫原： Synthesized peptide derived…

Experimental animals The generation of mice with tamoxifen-inducible, Tie2.Cre-ERT2-mediated deletion of LepR in endothelial cells was described previously12,17. For Cre recombinase activation, mice (6 weeks-of-age) were fed tamoxifen citrate-containing rodent chow (Envigo; TD.130860) for 6 weeks49. Genomic DNA from the brain, lung, small intestine, subcutaneous adipose tissue (SCAT) and visceral adipose tissue…

Author Contributions Conceptualization, L.W., W.B. and V.G.; methodology, Y.H., J.S. (John Sienkiewicz), J.S. (Jinying Sun), H.Z., H.H., S.X., R.B. and L.H.; software, Y.H., J.S. (John Sienkiewicz) and J.S. (Jinying Sun); validation, H.Z. and J.S. (John Sienkiewicz); formal analysis, V.G.; investigation, V.G. and L.W.; resources, L.W.; data curation, Y.H., J.S. (John…

Leukemia cell culture The human acute promyelocytic leukemia cell line (NB4) and the human acute lymphocytic leukemia cell line (MOLT-4) were purchased from Cell Lines Service (Eppelheim, Germany). Leukemia cells were cultured in RPMI-1640 medium supplemented with 10% (v/v) FBS and 1% (v/v) penicillin–streptomycin (Gibco Life Technologies, Waltham, MA, USA)….

Summary The Human Genome Sequencing Center (HGSC) provides a unique opportunity to gain high-throughput data production experience in an academic research environment, with opportunities for publication, conference travel and international scientific collaboration. As a member of the NGSI Clinical Informatics team, you will directly contribute to the HGSC’s pioneering work…

Study design The overall goal of this study was to explore the molecular mechanisms of remdesivir-related cardiotoxicity in anti-COVID-19 therapy. We first performed the GPCR screening using major anti-COVID-19 drugs as ligands (n = 3 per GPCR). We validated the results of GPCR screening by concentration-response analysis and determined the EC50 and…

Ethics statement All experiments with hamsters were performed in accordance with the Science Council of Japan’s Guidelines for the Proper Conduct of Animal Experiments. The protocols were approved by the Institutional Animal Care and Use Committee of National University Corporation Hokkaido University (approval ID: 20-0123 and 20-0060). All protocols involving…

Cell culture The MDA-MB231 (MB231) human breast cancer cell line was obtained from the American Type Culture Collection (ATCC, Manassas, VA) and grown in RPM1-1640 (Invitrogen) supplemented with 10% fetal bovine serum (FBS; Invitrogen, Waltham, MA) at 37 °C in a humidified atmosphere of 5% CO2 in the air. Cells were…

Balance equation of any chemical reaction online for freeCa3P2(s) + H2O(l) = Ca(OH)2(aq) + PH3(g) =>Ca3P2(s) + 6 H2O(l) = 3 Ca(OH)2(aq) + 2 PH3(g) Na + O2 = Na2O =>4 Na + O2 = 2 Na2O H2SO4 + NaOH = Na2SO4 + H2O =>H2SO4 + 2 NaOH = Na2SO4…

Cells and cell culture KOPN30, BV173, and K562 are BCR/ABL1-positive leukemia cell lines. All leukemia cell lines, as well as Ba/F3 cells, were maintained in RPMI-1640 medium supplemented with 15% fetal bovine serum (FBS) and penicillin–streptomycin (100 U/mL) at 37 °C in an atmosphere containing 5% CO2. KOPN30 cells were obtained…

Identification of MDV presence using open access sequencing data Metagenomic next-generation sequencing (NGS) data for field mosquitoes in China were searched from the NCBI SRA database (www.ncbi.nlm.nih.gov/sra) and CNGB Sequence Archive database (db.cngb.org/cnsa/) using the keywords “mosquito OR Anopheles OR Aedes OR Culex”. Followed by removing the datasets with less…

Materials BIS-Tris propane (BTP), glycine (Gly), sodium dodecyl sulfate (SDS), phosphate-buffered saline (PBS), bovine serum albumin (BSA), sodium chloride, Tween 20, and tris (hydroxymethyl) aminomethane (Tris) were obtained from Merck (Darmstadt, Germany). Sodium hydroxide was purchased from Avantor (Gliwice, Poland). All chemicals were of analytical grade. Deionized water was obtained…

Figure 1 shows the detailed workflow of the study. Fig. 1 General overview of the study. Briefly, MPXV was isolated from a skin lesion and then was used to infect CV-1 cells. After the designated infection times, total RNA was isolated and sequenced using direct cDNA sequencing protocol on ONT’s MinION platform….

1. Introduction Peptides are employed for a multitude of therapeutic approaches and can be obtained from the native organ or tissue in which they are produced as well as through recombinant methods where nucleic acids encoding a candidate gene(s) are delivered to and expressed in cells in vitro. As an…

Plasmid design The information on each plasmid for combinatorial libraries is indicated in Supplementary Table S1. Plasmids used in this study to generate combinatorial libraries for SpCas9, enAsCas12a, CHyMErA, enAsCas12a (dual-gRNA), CHyMErA.v2, multiSPAS were deposited to Addgene (IDs: 189632, 189633, 189634, 189635, 189636, 189637, respectively). 3Cs oligonucleotide design The protocol…

Ethics statement All experiments in this study comply with guidelines of the U.S. Public Health Service and were approved by the Institutional Biosafety Committee at the University of Pittsburgh. All animal husbandry and experimental procedures were conducted in accordance with U.S. Public Health Service policy and approved by the Institutional…

This paper describes an open-source digital image correlation algorithm for measuring local 2D tissue strains within tendon explants. The accuracy of the technique has been validated using multiple techniques, and it is available for public use. This protocol enables the measurements of the local tissue strains within a tendon during…

The proposed protocol includes guidelines on how to avoid contamination with endotoxin during the isolation of extracellular vesicles from cell culture supernatants, and how to properly evaluate them. This protocol provide guidelines on how to avoid contamination with endotoxin during the isolation of extracellular vesicles from cell culture supernatants, and…

Here, we present a protocol to isolate murine intestinal mesenchyme, including telocytes. These can be used for several applications, such as co-culture with mouse or human-derived organoids, to support growth and better reflect the situation in the original tissue. Our protocol for isolating intestinal mesenchyme results in a high yield…

The need to make quick therapeutic decisions requires procedures that allow reliable results to be obtained quickly and unambiguously. Such an ideal genetic test for detecting predictive mutations, i.e. those that already have a defined therapeutic path, should be characterized above all by the reliability of their results, simplicity, a…

Introduction Hyperuricemia is a disease caused by irregular purine metabolism, which is always accompanied by excessive production or abnormal excretion of uric acid (UA).1 Most notably, the overall incidence of hyperuricemia in China has been increasing over the past decade with the westernization of eating habits.2,3 However, there are blind…

5-Azacytidine pretreatment resensitizes cisplatin-resistant DLBCL cell lines to cisplatin to varying degrees A platinum-based anti-neoplastic agent is an essential component of salvage regimens for aggressive lymphomas, for example, rituximab, gemcitabine, dexamethasone, and cisplatin (R-GDP) for relapsed/refractory DLBCL and Hodgkin lymphoma [21]. As a single agent in humans, cisplatin is administered…

Preparation of GPMV GPMVs were prepared using different cell lines. Cells were cultured as follows: A549 (ATCC, Manassas, VA), HEK293 EBNA (herein referred to as HEK293, ATCC), HepG2 (ATCC), and Huh7 (ATCC) cells were kept in Dulbecco’s Modified Eagle’s Medium with high glucose (Sigma-Aldrich, Buchs, Switzerland) supplemented with 10% fetal…

Recombinant protein expression and purification NanoLuc is the tradename of an engineered deep-sea shrimp luciferase variant developed by Promega Corporation36. Genes encoding fusion proteins were inserted into pET-21d(+) vectors between NcoI and XhoI sites. IDO-Gal3 genetic and protein sequences are provided in Supplementary Information. Plasmids confer ampicillin resistance and were…

We reasoned that an ideal screening system for functional mRNA delivery would be model independent so that it could be applied in any preclinical model of disease, would consist of multiple measures of protein production that are each orthogonal to any others such that multiple formulations could be tested within…

Mice Trex1+/– mice (C57BL/6) were gifts from D. Barnes and T. Lindahl (Cancer Research UK). All animal experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and with the approval of the Institutional Animal Care and Use Committee (IACUC-DWZX-2020-511)….

Cell strains E. coli DH5α and B. subtilis 168 strains were grown overnight in Luria-Bertani broth at 37 °C and 180 rpm to a final OD600 ~1.7. B. subtilis strain 168 was kindly provided by Dr. S.A. Dubiley (Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia). T. adornatum strain…

Cell clusters in ZIKV-infected mouse testis defined by scRNA-Seq To investigate the influence of ZIKV infection on testes, testicular cells from ZIKV-infected (14 dpi.) and uninfected A6 male mice (Ifnar−/− mice) were analyzed by single-cell RNA sequencing (scRNA-Seq). After filtering out poor-quality cells, 11014 cells in control testes and 11974…

Plasmids LentiCRISPRv2 (Addgene; 52961) was purchased from Addgene (Cambridge, MA, USA). For HTT gene suppression, LentiCRISPR v2 plasmid with U6 promoter driving sgRNA expression and EF-1 alpha promoter driving the expression of Puromycin-T2A-HA-NLS-dCas9-NLS (or Puromycin-T2A-Flag-NLS-Cas9-NLS) was used. The sgRNA-Cas9 and sgRNA-dCas9 constructs were generated by cloning the CAG repeat-targeting sgRNA…

All methods were performed in accordance with institutional guidelines and regulations. CRISPR-Cas9 protein and guide RNA reagents All Cas9 proteins used in this study were derived from the prototypic Streptococcus pyogenes species. Cas9 protein was produced as described in Seamon et al.33 and contained a N-terminal nuclear localization signal and…

Introduction The metabolome is an incredibly diverse collection of small molecules (<1,500 Da) in biological systems involved in virtually every cellular process, including cellular energy production, macromolecule synthesis, epigenetic modifications, cell signalling and more (for recent reviews see [Citation1–6]). It responds rapidly (in seconds) to both internal (signalling, allostery) and external…

Acquiring amino acid substitutions via language model consensus We select amino acid substitutions recommended by a consensus of language models. We take as input a single wild-type sequence x = (x1,…,xN)∈ \(\mathcal{X}\)N, where \(\mathcal{X}\) is the set of amino acids, and N is the sequence length. We also require a set of…

Human CD34+ cell isolation Human fetal liver (FL) samples were obtained from Advanced Biosciences Resources (Alameda, CA) with proper consent. FL samples were cut in small fragments, treated for 25 min at 37 °C with Collagenase D (100 ng/mL; Roche). The cell suspension was prepared, and the human CD34+ cells were separated by…

Here we present a non-genetic method to generate human autologous liver spheroids using mononuclear cells isolated from steady-state peripheral blood. Generation of human liver spheroids using autologous sources can contribute to various research area like toxicology, cancer, and drug discovery. The main advantage of this technique is to use BDPC-derived…

Here, we describe an immunofluorescence-based method to quantify the levels of single-stranded DNA in cells. This efficient and reproducible method can be utilized to examine replication stress, a common feature in several ovarian cancers. Additionally, this assay is compatible with an automated analysis pipeline, which further increases its efficiency. Our…

The present protocol describes methods for evaluating DNA damage repair proteins in patient-derived ovarian cancer organoids. Included here are comprehensive plating and staining methods, as well as detailed, objective quantification procedures. The study of the DNA damage response is necessary in understanding the pathophysiology of ovarian cancer potential biomarkers in…

Ethics Human blood was obtained from healthy donors, approved by the Regional Ethics Committee and Imperial College Healthcare NHS Trust Tissue Bank (Regional Ethics Committee approval no. 17/WA/0161, Imperial College Healthcare Tissue Bank Human Tissue Authority license no. 12275, and Imperial College Research Ethics Committee no. 19IC5166). All samples were…

Constructs MOTH domains of human TALI (23-123), Otoraplin (18-128), and MIA (19-131), and TANGO1 (30-139) from Drosophila melanogaster (dmTANGO1(30-139)) were expressed from codon-optimized sequences in a modified pQE40 expression vector36 in M15 pRep4 E. coli strain. Human TANGO1 (21-131) (hsTANGO1 (21-131)) and TANGO1 (21-151) (hsTANGO1 (21-151)) were expressed from codon-optimized…

ENCODE ChIP-seq data analysis and prioritization of TAD boundary deletion loci Previously published chromosome conformation capture data was used for combined analyses and selection of TAD boundary deletion loci in this study, wherein TAD boundary calls were based on the maximum enrichment of CTCF at the TAD borders and their…

Cell culture and reagents Non-cancerous mammary epithelium cell (MCF10A), breast cancer cells including luminal (MCF7, MDA-MB-361, T47D, and BT474), HER2amp (SKBR3), and triple-negative (MDA-MB-231, CAL51, BT20, and MDA-MB-468) subtypes, as well as human embryonic kidney 293T (HEK-293T) cells, were purchased from the American Type Culture Collection (Manassas, VA, USA). MDA-MB-231,…

Cell culture and reagents HEK293 and HeLa cells (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal calf serum, 2 mM GlutaMAX‐1, 100 μg/mL streptomycin and 100 U/mL penicillin (complete growth medium) at 37 °C under 5% CO2. HeLa cells stably expressing GFP-TIFA were previously described8. Cells…

Study population and cell isolation PBMCs were obtained from leukocyte reduction chambers of 57 young (20–35 years old) and 64 older (65–85 years old) healthy blood donors of both sexes, purchased from the Stanford Blood Center and the Mayo Clinic Blood Center. The study was conducted in accordance with the…

IgG plasmids Antibody sequences and Fc-tagged ACE2 were cloned into the CMV/R plasmid backbone for expression under a CMV promoter. The antibodies with variable HC/LC were cloned between the CMV promoter and the bGH poly(A) signal sequence of the CMV/R plasmid to facilitate improved protein expression. The variable region was…

Gene Expression DataAssay Details Assay Reference: J:45678 Henderson DJ, et al., Over-expression of the chondroitin sulphate proteoglycan versican is associated with defective neural crest migration in the Pax3 mutant mouse (splotch). Mech Dev. 1997 Dec;69(1-2):39-51 Assay type: RNA in situ MGI Accession ID: MGI:5811768 Gene symbol: Pax3 Gene name: paired…

The present protocol shows how to differentiate CD3−/CD45+CD56+ cells with mild cytotoxicity from human expanded potential stem cells (hEPSCs) under both 3D and 2D culture conditions. This allows for routine phenotypical validation without the destruction of the complex microenvironment. Speaking of significance, generating NK cells from an expandable source is…

The present protocol describes two methods to measure caspase activity through a fluorogenic substrate using flow cytometry or a spectrofluorometer. This protocol allows researchers to identify if caspase activity occurs in conjunction with cell death, thus identifying different modes of cell death. The advantage of this technique is the flexibility…

To quantify the expression of different antigens, cells were stained using monoclonal antibodies coupled to phycoerythrin (PE), after which the number of PE molecules per cell was determined using quantitative flow cytometry. Although the number of PE molecules will not be exactly equal to the number of antigens, in this…

To maximize reproducibility, a list of the reagents and resources used in this study, as well as their source and identifier, is provided in Supplementary Table 5. Experimental model and subject details Oral swab samples were obtained from bottlenose dolphins (Tursiops truncatus) managed by the U.S. Navy MMP Biosciences Division, Space…

Introduction Cholangiocarcinoma (CC) is a form of gastrointestinal cancer that originates from the epithelium of either intrahepatic or extrahepatic bile ducts. It accounts for approximately 3% of all gastrointestinal cancers, with reported incidence of one to two cases per 100,000 persons per year in the USA (and much higher incidence…

Cells Human cervix epitheloid carcinoma HeLa cells (ECACC 93021013; RRID:CVCL_0030) were cultured at 37 °C in Dulbecco′s Modified Eagle′s Medium (DMEM) with 3 mM glutamine, 10% (v/v) fetal calf serum (FCS), 10 mM HEPES, 100 IU/ml penicillin, 100 μg/ml streptomycin, and 5 mM glucose in humidified air with 5% CO2. Human osteosarcoma 143B cells were purchased…

Reagents Activated Factor X (FXa) was purchased from New England Biolabs. Rivaroxaban was purchased from Santa Cruz. For Western blotting and immunofluorescence staining, the following antibodies were used: LOX rabbit polyclonal (ThermoFisher PA1-46020), LOXL2 rabbit monoclonal C-terminal (Abcam ab179810), LOXL2 polyclonal (Abcam ab197779), COLI monoclonal antibody (Invitrogen MA1-26771), COLIV rabbit…

OTUB1 deficiency causes defects in osteogenesis To determine the physiological role of OTUB1 in bone homeostasis, Otub1 global knockout mice (referred to as Otub1−/−) were constructed, and their phenotypes were analyzed on embryonic (E) day 14.5 (E14.5), E16.5, E18.5, and postnatal (P) day P0 (Fig. 1a and Supplementary Fig. 1a,…

Isolation of dental pulp stem cells Human pulp cells were extracted from healthy premolars of 25-year-old female (3U) and 19-year-old male (5C) who visited Kyushu University hospital for orthodontic treatment, with informed consent as described previously29. We used CD146-positive pulp cells as DPSCs because CD146 is thought to be a…

1. Introduction Long non-coding RNAs are identified as a type of non-coding RNA that is longer than 200 nucleotides, and which does not have the potential to be translated into protein. Long non-coding RNAs influence various cellular regulatory processes, including chromatin remodeling, RNA processing, transcriptional regulation, and translational control. Recent…

Plasmids The mingag fusion protein was generated by fusing a codon-optimised HIV-1 subtype B gp41 miniprotein (“min”) containing a fragment of the HR2 domain, the membrane proximal external region (MPER) and the transmembrane domain (TM) (HXB2: 8057-8345 bp)36 with a codon-optimised HIV-1 subtype B gagHXB2. A flexible GGGS linker was inserted…

Now rest is processed, and we follow save the files and move small to downstream analysis. Most genes in genomic repositories, genomics core sequence reads and genome and data? Pbs before you to focus on our health and quality controlled experiment or at the simulated ercc is due to systematic…

Purification of T7Select 10-3b phages The T7Select 10-3b phage vector (Novagen, Merck) was used in this study. Non-recombinant/recombinant T7Select 10-3b phages were purified from the lysate of infected E. coli strain BL5403 by PEG-8000 precipitation as per the method described in T7 select system manual. The pellet was suspended in…

B16 melanin(+) tdTomato cell line generation The generation and characterization of a lentivirus encoding tdTomato has been described previously22. The B16-D5-HER2 stable cell line was a generous gift from Louis Weiner (Georgetown University)13,23. Cell lines were cultured in DMEM Dulbecco’s modified Eagle’s medium (DMEM; Sigma) supplemented with 10% (v/v) fetal…

Ethics statement In vitro and animal work were conducted under appropriate biosafety conditions in a BSL-3 facility at the Institut für Virologie, Freie Universität Berlin, Germany. All animal experiments were performed in compliance with relevant institutional, national and international guidelines for the care and humane use of animals and approved…

Cell culture HEK293T, MEF-1, and U87 cell lines were used in this research. HEK293T was purchased from ATCC. MEF-1, and U87 cell lines were gifts from Muredach P. Reilly, Peter A. Sims, respectively. HEK293T, MEF-1, and U87 cells were cultured in DMEM with 4.5 g/L D-Glucose, supplemented with 10% fetal bovine…

Protein expression, purification and labeling The gene encoding the human A2AAR (UniProt C9JQD8) C-terminally truncated after residue Ala 316 (Supplementary Fig. 1a) was synthesized de novo (Eurofins). The nucleotide sequence was optimized for Leishmania tarentolae expression with the GeneOptimizer software (ThermoFisher Scientific). KpnI restriction site was introduced at the C-terminus and…

Errors after running jobscript for cellranger vdj pipeline 0 Hello, Below is pasted my jobscript to be submitted to run cellranger vdj pipeline on my fasta sample files. 1 #!/bin/bash 2 #PBS -N RED_aregvdj 3 #PBS -q serial 4 #PBS -l nodes=1:ppn=24 5 #PBS -l walltime=03:00:00 6 #PBS -l pmem=3300mb…

Plasmid construction The PVCpnf structural and accessory region (pvc1-16) and payload and regulatory region (Pdp1, Pnf and regulatory genes PAU_RS16570-RS24015) were synthesized de novo (GenScript) and cloned into pAWP78 and pBR322 backbones, respectively. All manipulations involving payload and regulatory plasmids (pPayload) involved standard PCR amplification with Phusion Flash 2x Master…

Research reported in this study complies with all relevant ethical regulations, and our animal study protocol has been approved by the Animal Experimental Board of Finland (license number ESAVI/28687/2020. Phage panning Sherpabody phage display library (size ~1011 cfu) was obtained from Next Biomed Therapies Oy. To develop sherpabodies specific for SARS-CoV-2…

1. Introduction Cancer is one of the main causes of death around the world, with the most significant being lung, liver, stomach, colon, and breast cancers. As a major concern, the dissemination of metastases is responsible for more than 90% of deaths by cancer. Tumor cells spread by lymphatic and…

C. elegans strains All strains were cultured under standard conditions78 and were always incubated at 20 °C during the experiments. The strains used were N2 (Bristol; WT): DREAM: MT8839 lin-52(n771) III, MT10430 lin-35(n745) I, MT15107 lin-53(n3368) I/hT2 [bli-4(e937) let-?(q782) qIs48] (I;III), MT8879 dpl-1(n2994) II, MT11147 dpl-1(n3643) II, JJ1549 efl-1(se1) V, BJS634…

货号： CSB-PA000018 规格： 图片： 其他： 产品详情 Uniprot No.： 基因名： CASP14 别名： Apoptosis related cysteine protease antibody; CASP 14 antibody; CASP-14…

The CHO cell line is an adherent epithelial cell derived from the ovary of Chinese hamster. It is widely used in biology and medical research of genetics, toxicity screening, nutrition, and gene expression, particularly to express recombinant proteins. CHO cells are the most common mammalian hosts for industrial production of…

doing PBS(population branch statistics) in ANGSD 0 Hi everyone, i am trying to do a Population branch statistic on my data i have done the first step where i calculate saf, now i need to do the second step calculating 2dsfs. when i submit the script on the cluster. I…

RevGel-seq sample preparation workflow Experiments were performed with the RevGel-seq protocol, capable of analyzing 10,000 input cells per sample with the specially designed gelation device (Fig. S3). The individual steps shown in Fig. 1A, from cell-barcoded bead coupling to library preparation for sequencing, are more fully described below: Cell labeling Cells…

Experimental animals and DOX treatment All animal experiments performed on live animals were approved by Fudan University’s Committee on the Ethics of Animal Experiments and adhered to relevant guidelines including the ARRIVE guidelines for animal experiments in the study. Adult male C57BL/6 mice (8–10 weeks old; 23–26 g; Department of Laboratory…

This protocol will allow researchers to not only streamline their analysis pipeline, but it will also allow them to extract more information out of their organoid stream, thereby enhancing the translational relevance. The main advantage of this protocol is it combines a streamlined workflow with an automated and clinically relevant…

Reagents, consumables, and kits The oligonucleotides used as primers in the asymmetric polymerase chain reaction (aPCR) and as staple strands for the folding of PB NPs, along with the CpG ODN 1018 adjuvant were purchased from Integrated DNA Technologies (IDT). Fluorescently labelled oligonucleotides used in FRET assay, FAM (donor) and…

Experimental design The objective of this study was to test the hypothesis that CAR can improve cardiac function after MI by containing the lesion and improving remodeling. We used an in vivo loss-of-function approach with coronary artery ligation in inducible heart-specific CAR knockout mice. In a complementary gain-of-function approach, we…

Hello, I have tried running mrbayes-v3.2.6 on my university cluster with a text block in my nexus file, as below: begin mrbayes; set autoclose=yes nowarn=yes; CHARSET mtgenome = 1-16701; partition favored = 1: mtgenome; set partition = favored; unlink shape=(all) pinvar=(all) statefreq=(all) revmat=(all) tratio=(all); prset applyto=(all) ratepr=variable; lset applyto=(1) nst=2…

“The World Health Organization on Friday called on China to release new data linking the Covid pandemic’s origins to animal samples at Wuhan Market after the country recently took down the research,” reports CNBC. The existence of the new data was revealed by the Atlantic earlier this week, in an…