Tag: FACS

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)…

About Serotiny Serotiny is a company that is engaged in the discovery of proteins for cell therapies. It focuses on such proteins as CARs (chimeric antigen receptors) for the treatment of cancers and genetic disorders. Job Description Description Serotiny invents multi-domain proteins for gene and cell therapies including genome editors…

Significance The collection of large amounts of whole-genome sequencing data allowed for identification of mutational signatures, which are characteristic combinations of substitutions in the context of neighboring bases. The clinical significance of these mutational signatures is still largely unknown. In neuroblastoma, we showed that high levels of cytosine > adenine…

September 1, 2021 | Harvard-Israel and Sema4-Avera launch precision medicine partnerships, Rice announces new Genetic Design and Engineering Center, Foundation Medicine partners with Epic, and Amgen and USC provide cryo-TEMs. Plus new products from BioIVT, BD.   Pluto Biosciences has launched its cloud-based, collaborative life sciences platform that leverages technology…