Tag: FACS

In a recent study published in the Proceedings of the National Academy of Sciences, researchers developed a fully synthetic platform for nanobody generation. Study: Fully synthetic platform to rapidly generate tetravalent bispecific nanobody–based immunoglobulins. Image Credit: Adao / Shutterstock.com What are nanobodies? Biologics have been increasingly used for the treatment or prevention…

Dermal cell characterization by 10X scRNA-seq. a, Workflow used to obtain dermal cells of adult and aged mouse back skin. Single-cell suspensions were enriched separately for EpCAM–CD45– and CD45+ cells by FACS. Transcriptomes of sorted single cells were then analyzed by 10X scRNA-seq. For CD45+ cells, n = 7 mice for the adult…

Cell culture and cell line construction Cells were cultured at 37 °C in 5% CO2 atmosphere with 100% humidity. Telomerase-immortalized RPE-1 retinal pigment epithelium cells (CRL-4000, American Type Culture Collection), U2OS osteosarcoma cells (HTB-96, American Type Culture Collection) and derivative cell lines were grown in DMEM/F12 (1:1) medium without phenol red…

Natera%2C+Inc. (NASDAQ: NTRA), a global leader in cell-free DNA testing, today announced new data on its Prospera test being presented at the American Transplant Congress (ATC) 2023 taking place June 3-7, 2023. The presentations include several interim analyses from ProActive, a prospective donor-derived cell-free DNA (dd-cfDNA) study evaluating Natera’s Prospera…

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…

Single-cell and single-nucleus RNA sequencing of sarcoma subtypes In this work, we studied sarcomas from varying tissue origins, including osteosarcoma (OS), Ewing sarcoma (ES), and desmoplastic small round cell tumor (DSRCT) (Fig. 1). We used different dissociation protocols: Miltenyi Tumor Dissociation Kit, cold-active protease derived from Bacillus licheniformis, and Nuclei EZ…

An optimized CRISPR knock-in method using long dsDNA donors for efficient tagging with fluorescent proteins in human diploid cells. Given the widespread use of CRISPR-mediated generation of knock-in cell lines, it is informative to compare the performance of long dsDNA and ssDNA donors in a simple and practical knock-in method….

Revsbech, N. P. et al. Determination of ultra-low oxygen concentrations in oxygen minimum zones by the STOX sensor. Limnol. Oceanogr.: Methods. 7, 371-381. (2009). Wright, J. J., Konwar, K. M. & Hallam, S. J. Microbial ecology of expanding oxygen minimum zones. Nat. Rev. Microbiol. 10, 381–394 (2012). Article  CAS  PubMed …

Tucked away in basements, down long hallways, and in buildings scattered across campus sits billions of dollars in equipment, waiting to help researchers and students create breakthroughs in myriad fields. Postdoctoral scholar Arianne Caudal and research scientists Norah Brown, Soren Faulkner, and Garvey McKenzie using the Orbitrap Eclipse Tribrid mass…

Annotation of short-read scRNA-seq data with isoform-level information Using fluorescence-activated cell sorting (FACS), we isolated the Lineage-negative, cKit (Cd117) positive (LK) cell fraction of mouse bone marrow cells, a population containing stem and progenitor cells14 from young (8 weeks old, n = 3) and aged (72+ weeks old, n = 3) mice. We generated…

Who are we looking for? We are seeking a Flow Cytometry Facility Analyst/Manager to join School of Biomedical Sciences, Faculty of Health, Academic Division. The appointee will work in the Centre for Microbiome Research (CMR) performing fluorescence activated cell sorting (FACS) of microbial cells and viruses from a wide range…

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…

Screening strategy for identifying viral inhibitors of the cGAS/STING pathway Vaccinia virus is a large cytoplasmic DNA virus with a 190 kilobase pairs (kbp) genome that encodes over 200 proteins21,22. MVA has an approximately 30-kbp deletion from its parental vaccinia genome, resulting in the loss of many immune-modulatory viral genes15….

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…

When the Sloan Kettering Institute opened its doors on Friday, April 16, 1948, an Associated Press wire story announced: “The world’s greatest cancer center opened here today.” And over the last seven and a half decades, institute researchers have made important contributions to the fundamental understanding of human biology, as well as…

Nathan Steinle, MD: In early 2022 in the United States, faricima became available and at least first in my knowledge it came out for both DME [diabetic macular edema] and wet AMD [age-related macular degeneration] at the same time. Dr Scott, can you talk to me a little bit about…

During a plenary session at the American Urological Association’s 2023 Annual Scientific Meeting, Neal D. Shore, MD, FACS, Director, CPI, of Carolina Urologic Research Center, and a CMO with GenesisCare, US in Myrtle Beach, South Carolina, presented findings from the phase 3 EMBARK trial demonstrating a strong oncologic benefit of…

About the team/job The Hackett group studies the epigenetic basis of genome regulation across scales, from chromatin function to organism response. We aim to to understand the causal and context-specific impact of chromatin states in healthy and diseased tissues, and the potential for epigenetic memory. More information can be found…

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…

Phylogenetic tree construction Tree diagram showing relationships between CDK proteins was constructed from a multi-sequence alignment (MSA) using Geneious95. The “Geneious Aligner”, was used to generate the MSA, and the neighbor joining method was used to construct the tree. All default parameters were used except where otherwise indicated. Combinatorial CRISPR…

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…

CRISPR screen targeting E3 ligases identifies major regulators of Parkin-independent mitophagy A. Schematic depicting the CRISPR screening strategy. RPE1-Cas9i-mt-mKeima cells were transduced with a lentiviral CRISPR sgRNA library targeting 606 E3 ligases. sgRNA-expressing cells were selected for 7 days with puromycin (Puro) and Cas9 expression induced with doxycycline (Dox) for 9 days….

Robert L. Ferris, MD, PhD, FACS The primary end point of overall response rate (ORR) benefit was not met with first-line nivolumab (Opdivo) plus ipilimumab (Yervoy) vs nivolumab alone among patients with platinum-refractory recurrent or metastatic squamous cell carcinoma of the head and neck (R/M SCCHN) in the CheckMate 714…

Ethics declaration Animal study was performed following the Guide for the Care and Use of Laboratory Animals. Animal experiment was approved by the Institutional Animal Care and Use Committee of Colorado State University (protocol number 1090) in advance and conducted in compliance with the Association for the Assessment and Accreditation…

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…

Findings from the phase 3 EMBARK trial showed that enzalutamide (Xtandi) plus leuprolide reduced the risk of metastasis or death by nearly 60% vs placebo plus leuprolide in patients with nonmetastatic hormone-sensitive prostate cancer (nmHSPC) with high-risk biochemical recurrence (BCR).1,2 “The EMBARK study is a phase 3 trial exploring the potential of…

Future research efforts in the gynecologic cancer space should center on diversity in clinical trials, combined modality types, and surgical research, according to Robert L. Coleman, MD, FACOG, FACS. In an interview with ONCOLOGY®, Coleman discussed the future of gynecologic cancers and the importance of attending and engaging in meetings…

Abstract Muscular pathologies represent a class of diseases and disorders comprising both inherited genetic disorders (i.e. muscular dystrophies) and secondary conditions (i.e. cachexia). The most promising therapeutics for these conditions are adeno-associated virus (AAV)-mediated gene therapies. This strategy has shown promise in clinical trials; however, immunogenicity and lack of specificity…

Transcriptional activation of RCOR2 by neo-TADs in RELA ependymoma. a Chromatin contacts in a reconstructed ZFTA tumor genome (sample 4EP53) including the tandem duplication that leads to the ZFTA-RELA fusion (chr1:63532174-65429788, green boxes). Solid black boxes show TADs identified by applying TopDom to the Hi-C data mapped on to the…

Materials The materials used for cell cultures and characterization are listed in Additional file 1: Table S1. Cell sources After getting informed consent, PB was drawn from three healthy O, Rh D-positive donors. CB was collected from three healthy newborn babies at the Department of Obstetrics and Gynecology at Severance…

What is the standard of validating differentially expressed genes experimentally from scRNA seq analyses? 1 Are we required to carry out ddPCR to validate the DGEs obtained from scRNA seq analyses? singlecell scRNA validation • 38 views There is no standard if you ask me. I cannot recall a single-cell…

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…

Overview of the study To uncover the contexts and biological processes that affect gene expression regulation, this study took advantage of both the resolution of single-cell data and the directionality captured by co-eQTLs. First, we constructed cell-type-specific co-expression networks using five scRNA-seq PBMC datasets from three recently generated PBMC scRNA-seq…

– GENESIS trial achieved statistical significance (p<0.0001) across all primary and secondary endpoints – – Nature Medicine publication describes how GENESIS trial included patients representative of current multiple myeloma population undergoing autologous HSCT, including older patients and those who received lenalidomide-containing induction therapies – factors associated with impaired HSPC mobilization…

BioLineRx Ltd. (BLRX) today announced the publication of data from the Company’s GENESIS Phase 3 clinical trial in the peer-reviewed journal Nature Medicine. The international GENESIS trial evaluated the safety and efficacy of the Company’s lead investigational candidate motixafortide plus granulocyte colony-stimulating factor (G-CSF) versus placebo plus G-CSF for the…

Patient demographics were comparable across study cohorts From 22 January 2018 to 30 October 2020, a total of 122 patients from 18 sites in five countries were enrolled and randomized 2:1 to receive either motixafortide + G-CSF (80 patients) or placebo + G-CSF (42 patients) for HSPC mobilization (Fig. 1 and Extended Data Fig. 1a). Demographics between the two treatment…

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…

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…

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…

OCT4 alone was insufficient to reprogram PBMCs into iMSCs directly Previously, we reported that lentivirally expressed OCT4 could directly reprogram human cord blood CD34+ hematopoietic progenitor cells into iMSCs with very high efficiency11. Therefore, we first tried to convert human PBMCs into iMSCs by overexpressing OCT4 alone using a clinically…

To meet the challenge of sample depletion during preparation, there have been a number of recent advances in what is now a rapidly moving field of study of SCP, with important contributions from several investigators. Earlier work combined capillary electrophoresis (CE) with high-resolution mass spectrometry (HRMS) to analyze proteomes of…

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…

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…

Poxvirus vectors for directed evolution of GPCRs in mammalian cells Monoclonal and homogenous expression of the gene of interest are critical prerequisites for selections in directed evolution that are difficult to achieve in mammalian cells with common expression strategies such as transient transfection. To reach both equal gene dosage and…

Mice Rosa26-YFP48, Rosa-tDTomato49, K14creER, Lgr5creER (ref. 50), KrasLSLG12D (ref. 51) and Trp53fl/fl (ref. 52) mice were obtained from the NCI mouse repository and Jackson Laboratories. Rhojfl/fl mice53 were a gift from A. Uemura. All mice used in this study were composed of males and females with mixed genetic background. Mouse…

Knockdown of EWSR1::CREM and CREM expression in CHL-1 cell line induced senescence and reduced proliferation and migration Loss of EWSR1::CREM fusion protein in CHL-1 has previously been found to be associated with reduced proliferation, senescence and impaired invasion3. To validate these findings, we investigated the cellular properties associated with endogenous…

Job description This is a research support post in Mishto lab within the Centre for Inflammation Biology and Cancer Immunology (CIBCI) at the Guy’s campus of King’s College London. This post is also shared with Kordasti lab at the Comprehensive Cancer Centre at the Guy’s campus of King’s College London…

Characterization of klara. (A) Expression of csf1ra was analyzed via RT-PCR using cDNA from FACS-sorted populations of lymphocytes, progenitors, and myeloid cells obtained from the whole kidney marrow (WKM) of a wild-type N. furzeri. Csf1ra was detected in all subpopulations, most strongly in myeloid cells. As a negative control, an…

Transcribed image text: A. What type of FACS data graphical representations are Panel A and Panel B? (1 Mark) B. All data displayed in Panel B have been gated from data displayed in Panel A using a FSC and SSC gate. What does this statement mean? Why would you want…

Screening for hyper-potent shRNA against SARS-CoV-2 We parsed the SARS-CoV-2 genome into a series of potential short hairpin RNA (shRNA) targets (Supplemental Fig. 1). This process was conducted by tiling the genome with overlapping 50 nucleotide-long sequences, each shifted by a single nucleotide from the other. The region targeted by each…

ISC Class 12th Biotechnology Syllabus 2023: Biotechnology is a field of science that applies biological processes, organisms, or systems to develop products or technologies that improve human lives. It involves using living organisms or their derivatives to create new products or processes. Biotechnology has multiple applications in various areas such…

General Questions What information should I know before I do the RIP-Seq? Before doing a RIP-SEQ you generally need to specify • The type of sample to be done • What RBPs are of interest • Whether IP-level antibodies are available for these RBPs • The type…

Received: 24 January 2023 / Revised: 2 March 2023 / Accepted: 4 March 2023 / Published: 7 March 2023 Round 1 Reviewer 1 Report In this manuscript, Staneva et al. study mitochondrial dynamics of ageing in budding yeast chromatin mutants. In particular, the authors investigated arp4, hho1 and double mutants….

Variants in centrosome genes were associated with malignant peripheral nerve sheath and gastrointestinal stromal tumors, and heritable defects in the shelterin complex were linked with increased susceptibility for sarcoma, melanoma, and thyroid cancers, according to findings from the International Sarcoma Kindred Study (ISKS). For the study, investigators examined the whole-genome…

Patients and clinical data collection The institutional review board of the Faculty of Medicine, Chulalongkorn University approved this study (IRB No. 264/62) which follows the Declaration of Helsinki Guidelines and all subsequent amendments. Written informed consents were obtained from parents or legal guardians of the participants. From June 2016 to…

Lentiviral vector construction and production The lentiviral vector was designed as previously described [13, 41]. LdCV vector contains the following: human codon-optimized S. pyogenes dCas9 which was fused at the C-terminus with VP64-p65-Rta (VPR). LAT-TCS-dCas9-VPR was assembled by fusing LAT (Human cDNA, NM_001014987.2) with dCas9-VPR-Q8 and cloned into a modified…

Cell lines Immortalised HA1E (hTERT and SV40ER) and tumourigenic HA1ER cells (hTERT, SV40ER and HRAS-G12V) from stepwise tumourigenesis models generated from normal human embryonic kidney cells were obtained from Dr. Hahn (Broad Institute of MIT and Harvard, Cambridge, USA). Cells were cultured in DMEM (1 g/L glucose) (Gibco, cat. #10567014) supplemented…

Data reporting No statistical methods were used to predetermine sample size. The experiments were not randomized. The investigators were not blinded to allocation during experiments and outcome assessment. Study approval This study and all the relevant experiments were approved by Guangzhou Eighth People’s Hospital Ethics Committee (No. 202001134 and 202115202)….

Biochemical Sciences Location: Guildford Salary: £34,308 to £38,592 per annum Fixed Term Post Type: Full Time Closing Date: 23.59 hours BST on Sunday 26 March 2023 Reference: 007423 Primary work place: Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey GU2 7XH Line manager & Principal Investigator: Dr Patrizia Camelliti Applications are…

1 Introduction Recombinant therapeutic proteins, also known as biologics, have been successfully used to treat a wide range of diseases, including cancers, inflammatory disorders, and cardiovascular diseases (Walsh, 2018). Biosimilars with comparable efficacy and safety profiles have emerged as alternative options to innovator therapies. A biosimilar is a biologic that…

Introduction According to the global cancer statistics 2018, breast cancer has the highest global incidence (28.9%) and mortality rates (23.0%) in women.1 Chemotherapy, as a mainstay in clinical breast cancer treatment, is limited by severe side-effects resulting from the nonspecific drug distribution in the human body.2–4 What is worse, a…

If your news feed looks anything like ours, you’ve likely stumbled across concerning reports that seem to link breast implants and certain types of cancer. Most recently, in early September of 2022, the U.S. Food and Drug Administration (FDA) released a new report that states breast implants may be linked…

Ethics statement All animal experiments were performed in compliance with the Regulations for the Care and Use of Laboratory Animals issued by the Ministry of Science and Technology of the People’s Republic of China, which enforces the ethical use of animals. The protocol was approved by Institutional Animal Care and…

HOLLYWOOD, Fla. – Researchers from The Hebrew University of Jerusalem and their collaborators are harnessing cell-free DNA (cfDNA) methylation signals as potential biomarkers for cardiac diseases. Presenting preliminary results from his group in a Tuesday evening talk at the Advances in Genome Biology and Technology annual meeting held here this…

Start Current SNIC Small Compute Projects SNIC 2022/22-1097 Cancer metastasis via RNAseq in the chicken olfactory neurons SUPR uses JavaScript for certain functions. We cannot guarantee that you will be able to use the system with JavaScript disabled. Primary Classification: 30401: Medical Biotechnology (focus on Cell Biology (incl. Stem Cell…

Ethics statement All fresh peripheral blood samples were obtained after approval of protocols by the Weizmann Institutional Review Board (IRB application 92-1) and following informed consent from the donors. The study using BAL fluid samples was approved by the Hadassah Medical Organization research ethics committee in accordance with the Declaration…

Isolation of pure populations of MuSCs and niche cells by fluorescence-activated cell sorting (FACS) Hindlimb muscles were dissected from young (4–6 weeks old) or aged (22–26 months old) C57BL/6J (Jackson Laboratory, 000664) mice and minced until no visible tissue chunks were visible. Muscle was then digested in a 15 ml Falcon…

Single sample GSEA analysis 1 Hi, I sorted specific type of cells by FACS and subjected it to RNA-seq. I want to test the enrichment of target cell type. I want to perform analysis just like GSEA to show the enrichment and get the plot which is common for GSEA…

Here we provide a tool that directly selects biallelically edited knock-ins using cell-surface displays, thereby eliminating the need for extensive clonal verification. By doing so SNEAK PEEC greatly reduces the considerable effort currently expended to obtain biallelically edited clones. The ability to generate a clonal population with multiple edits is…

counting reads in a fastq file using a refrence fasta file 1 Hi I have a reference fasta file containing 130,000 unique sequences (barcodes), each 30nts long. These sequences were synthesized with random incorporation of nucleotides in each position, so they have very large hamming distance. I had a pool…

Category Research / Academic Location Groningen We are looking for a highly motivated and enthusiastic Postdoctoral researcher with strong backgrounds in chemistry, biochemistry, biotechnology or closely related fields to join the research group of Dr Sandy Schmidt, (schmidt-researchgroup.com) at the Department of Chemical and Pharmaceutical Biology (www.rug.nl/research/grip/cpb/) within the Groningen…

Abstract Circular RNAs have been reported to be widely involved in cancer cell tumorigenesis and drug resistance; here, the aim of this study was to investigate whether circRNA Integrin Subunit Alpha 7 (ITGA7) (circ_ITGA7) was associated with the tumor growth and radiosensitivity of colorectal cancer (CRC). We found that circ_ITGA7…

Abstract The ability to map genetic interactions has been essential for determining gene function and defining biological pathways. Therefore, a system to readily perform genome-wide genetic modifier screens in human cells is a powerful platform for dissecting complex processes in mammalian cells, where redundancy and adaptation commonly mask the phenotype…

Generation and validation of MUC5B/AC-KO cultures. (A) Overview of the MUC5B/MUC5AC gene-targeting approach via lentivirus-mediated delivery of sgRNA and CRISPR-Cas9. Transduced and sorted cells were differentiated at air-liquid interface (ALI). Once fully differentiated, the mucus gel was collected by apically washing cultures. HEK293T, human embryonic kidney–293T; FACS, fluorescence-activated cell sorting;…

Cells and viruses hACE2 expressing BHK-21 (BHK-21-hACE2) and Vero-E6 cells were provided by State Key Laboratory of Virology, Wuhan University. Vero-E6 cells were cultured in DMEM (Gibco, Cat.11995–040) supplemented with 10% fetal bovine serum (Gibco, Cat.10091–148) and 1% penicillin and streptomycin solution (BasalMedia, Cat.S110JV) at 37°C with 5% CO2. BHK-21-hACE2…

Secondary use of biopsies as research specimens The ten melanoma samples used in this study were collected by Sanford Health and classified by a board-certified pathologist. Their secondary use as research specimens (fully de-identified formalin-fixed, paraffin-embedded (FFPE) blocks) was approved under Sanford Health IRB protocol 640 (titled ‘Understanding and improving…

Ethics statement All procedures involving laboratory mice were approved by the Duke University IACUC under the protocol numbers A189-18-08 and A142-21-07. Mice were housed with up to five mice per cage and the ambient room conditions ranged from 70–74 °F and 30–70% humidity with a 12-hour dark/light cycle. Animals were assessed…

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causal agent of the ongoing coronavirus disease 2019 (COVID-19) pandemic, has claimed more than 6.5 million lives worldwide. The genomic evolution of the virus has led to the rapid emergence of new SARS-CoV-2 strains, and several of these variants are more infectious and…

In a recent study posted to the bioRxiv* preprint server, researchers developed a three-dimensional (3D) in vitro tissue model to assess the effects of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on the vascular endothelial cells. Study: SARS-CoV-2 spike protein induces endothelial dysfunction in 3D engineered vascular networks. Image Credit:…

Experimental methods Stem cell and organoid culture We used six human iPS cell lines (Hoik1, Wibj2, Kucg2 from the HipSci resource47; 409B2 from the RIKEN BRC cell bank; 01F49i-N-B7 (B7) from Institute of Molecular and Clinical Ophthalmology Basel; and WTC from the Allen Institute) and three human ES cell lines (H1-PAX6YFP…

Reagents, antibodies, viruses and cells LMW-Poly(I:C), LPS, 2′3′-cGAMP and DMXAA (InvivoGen); hydroxyurea, camptothecin and mitomycin C (MCE); GM-CSF, Flt3L (peproTech); lipofectamine 2000 (Invitrogen); polybrene (Millipore); RNAiso Plus (Takara); HT-DNA (Sigma); SYBR (BIO-RAD); dual-specific luciferase assay kit (Promega); ELISA kit for murine Ifn-β (PBL); ELISA kits for murine IP-10 (Biolegend) were…

This protocol describes the induction of Gram-negative monobacterial sepsis in a mouse model system. The model is useful in investigating the inflammatory and lethal host responses during sepsis. Sepsis is a dysregulated host immune response to microbial invasion or tissue damage, leading to organ injury at a site distant from…

scRNAseq replicates. CD45.1+ P14 cells were adoptively transferred into adult CD45.2+ GP or GFAP:GP mice 24h before i.c. infection with rLCMV-GP33. At least six weeks later, mice were treated with antiCD8a depleting antibody or isotype control. One week after depletion, mice were treated with TAM i.p. for four consecutive days…

Introduction The clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) protein systems naturally exist in prokaryotes. They are RNA-mediated defense mechanisms to protect bacteria and archaea from invading nucleic acids (Barrangou et al., 2007). CRISPR–Cas systems are classified into two classes (1 and 2). Class 2 CRISPR–Cas require a single…

Website Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) Job Category: Bioinformatics Job Job ID: 22/16 Date Posted: 05.05.2022 Deadline: 19.06.2022 Location: Jena, Germany The Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) in Jena is a federal and state government-funded research institute and member…

Single sorted bone marrow-derived CD45+ EpCAM+ cells express both the CD45 and EpCAM genes and surface proteins. CD45+ EpCAM+ cells were sorted to high purity and analyzed for CD45 and EpCAM expression using immunofluorescence microscopy, as well as qRT-PCR. CD45+ EpCAM- cells were also sorted as a control. (a) FACS…

Coronavirus disease 2019 (COVID-19) vaccines have played a critical role in reducing transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission. However, with emerging reports of waning vaccine efficacy, there remains an urgent need to develop prophylactic measures against COVID-19. In a recent study published on the bioRxiv*…

Hi, in this video we’ll show you how to explore the m6A and m5C epitranscriptomes upon viral infection. To start the experiment, you’ll need 50 million cells for the noninfected and 50 million for the infected condition. In order to obtain a high quality dataset upon analysis, you’ll need to…

Name Rat Anti-Mouse CD25/IL-2R Antibody (GWB-31929F) Related Product Names Rat Anti-Mouse CD25/IL-2Ra-PE; PMP-22; Protein CD25; SR13 myelin protein; Schwann cell membrane glycoprotein; SAG Cd25; Pmp-22Rat Anti-Mouse CD25/IL-2RPmp22 Gene Symbol IL2RA Gene Fullname Interleukin-2 receptor subunit alpha Gene id 3559 Swissprot ID P01589 Protein Name Interleukin-2 receptor subunit alpha Protein Accession…

Cell lines and culture Unless stated otherwise, all tumor target cell lines were sourced from the ATCC via LGC Standards. Human endogenous mesothelin (MSLN)-positive target cell lines H-226 (lung carcinoma, MSLN+, ATCC® CRL-5826™) and AsPC-1 (ATCC CRL-1682), and MSLN-negative Raji control cells (ATCC CCL-86) were cultured in RPMI-1640 Glutamax (Life Technologies,…

Integration of APC scRNA-seq data reveals heterogeneity of adipocyte progenitor cells In a previous study9, we defined Lin−Sca1+CD142+ APCs as adipogenesis regulatory (Areg) cells and demonstrated that these cells are both refractory toward adipogenesis and control adipocyte formation of APCs through paracrine signaling. In contrast, Merrick et. al.4 observed that Lin−CD142+ cells…

Significance The plant-specific H3K27me1 methyltransferases ATXR5 and ATXR6 play integral roles connecting epigenetic silencing with genomic stability. However, how H3K27me1 relates to these processes is poorly understood. In this study, we performed a comprehensive transcriptome analysis of tissue- and ploidy-specific expression in a hypomorphic atxr5/6 mutant and revealed that the…

Significance Interferon (IFN)-mediated antiviral responses serve as the first line of the host innate immune defense against viral infection. Here we identify a previously uncharacterized protein designated phospholipase A2 inhibitor and LY6/PLAUR domain-containing protein (PINLYP), which is essential for embryonic development and plays an important role in type I IFN…

Starting in summer 2021, we ran 6 rounds of IL-2R binder design puzzles. Now we’ve selected 115 promising Foldit designs to test for binding in the lab! In early 2022, we will conduct a FACS experiment to test if these protein designs successfully bind to the IL-2R target. Targeting IL-2R…

STICR lentiviral library preparation and validation We synthesized a high-complexity lentivirus barcode library that encodes approximately 60–70 million distinct oligonucleotide RNA sequences (STICR barcodes). STICR barcodes comprised three distinct oligonucleotide fragments cloned sequentially into a multicloning site within the 3′ UTR of an enhanced green fluorescent protein (eGFP) transgene under…

BD Influx Sorter, a globally unique prototype equipped with fiber optics and octagon/trigon arrays. BD Influx Sorter, a globally unique prototype equipped with fiber optics and octagon/trigon arrays. Flow Cytometry Fluorescence-Activated Cell Sorting (FACS) is a technology that enables qualitative and quantitative analysis of cell mixtures and sorting by individual…

Introduction Pancreatic ductal adenocarcinoma (PDAC) is a lethal gastrointestinal disease with increasing morbidity, which also has a growing impact on cancer-specific mortality worldwide.1 Nearly 40% of all PDAC cases are localized to the pancreas and characterized with the involvement of major vascular structures, leading to unresectable disease without metastases detected…

Large-scale initiative will advance innovative research, improved health outcomes CHARLESTON, S.C. and SAN MATEO, Calif., (Sept. 20, 2021) – The Medical University of South Carolina  (MUSC) and Helix have announced a strategic collaboration to develop a first-of-its-kind population genomics initiative in South Carolina called In Our DNA SC. The large-scale program is designed…

The mammalian genome is characterized by widespread methylation of cytosine residues. After fertilization, however, both maternal and paternal genomes undergo extensive demethylation, reaching a low point in the blastocyst (1⇓⇓–4). The embryo genome is then remethylated by the activity of de novo DNA methylation enzymes (5). Mouse embryonic stem (ES)…