Journal: Cell

Article Title: Cannabinoid receptor 1 antagonist genistein attenuates marijuana-induced vascular inflammation

doi: 10.1016/j.cell.2022.04.005

Figure Lengend Snippet: (A) The effects of Δ9-THC on cell viability of human coronary artery endothelial cells (HCAECs), human umbilical vein endothelial cells (HUVECs), normal human cardiac fibroblasts-ventricular (NHCF-V), and human embryonic stem cell-derived cardiomyocytes (hESC-CMs). Cells were treated with increasing concentrations of Δ9-THC for 48 h, and cell viability was measured by the CellTiter-Glo luminescent cell viability assay.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) , ATCC , CRL-1730.

Techniques: Derivative Assay, Cell Viability Assay

Journal: Cell

Article Title: Cannabinoid receptor 1 antagonist genistein attenuates marijuana-induced vascular inflammation

doi: 10.1016/j.cell.2022.04.005

Figure Lengend Snippet: Δ 9 -THC induces cytotoxicity in human endothelial cells

Article Snippet: Human umbilical vein endothelial cells (HUVEC) , ATCC , CRL-1730.

Techniques: Virus, Recombinant, Membrane, DNA Ligation, Luminex, RNA Sequencing, Knock-Out, Control, Plasmid Preparation, Expressing, Software, Modification, Diagnostic Assay

Journal: STAR Protocols

Article Title: Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells

doi: 10.1016/j.xpro.2020.100028

Figure Lengend Snippet: FACS Plots Showing Lin − Sca1 + c-Kit + (LSK) Cell Fractions after MACS Purification and 2 Days of Culture Bone marrow cells were stained with α-mouse lineage cocktail, Sca1, and c-Kit antibodies. (A) The negative fraction of the LS column should be almost completely depleted of Sca1 + cells. (B) The positive fraction eluted from the LS or MS separation column is enriched in LSK cells and progenitor cells, which all express Sca1. The LSK population should comprise 6 to 8% of the Sca1 + cell fraction. Lymphoid progenitor cells require IL-7 to survive and will be depleted upon culturing with 50 ng/mL mTPO, mSCF, mFLT-3, and hIL-11. (C) Purified LSK cell fraction after 2 days of culture in HSPC medium.

Article Snippet: Anti-mouse Sca1 MicroBead Kit (VioBrightTM FITC) , Miltenyi Biotec , Cat#130-123-124.

Techniques: Purification, Staining

Journal: STAR Protocols

Article Title: Protocol for Efficient CRISPR/Cas9/AAV-Mediated Homologous Recombination in Mouse Hematopoietic Stem and Progenitor Cells

doi: 10.1016/j.xpro.2020.100028

Figure Lengend Snippet:

Article Snippet: Anti-mouse Sca1 MicroBead Kit (VioBrightTM FITC) , Miltenyi Biotec , Cat#130-123-124.

Techniques: Virus, Recombinant, DNA Extraction, Cloning, Modification, Plasmid Preparation, Software, Adhesive, Real-time Polymerase Chain Reaction, Sterility, Centrifugation

Journal: Journal of Translational Medicine

Article Title: A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors

doi: 10.1186/1479-5876-11-23

Figure Lengend Snippet: Construction and characterization of the CD19-IgG 1 Fc fusion proteins. A . Diagrams of the lentiviral vector provirus constructs used to transduce the CD19sIg fusion genes. B . Reduced protein electrophoresis by SDS PAGE of fusion protein products after concentration and purification using Protein A Dynabeads. Centricon concentrated supernatant samples from fresh Pro293a™-CDM (A) , 293 T cells (B) , 293 T cells expressing CD19sIg1-3 (C) , and 293 T cells expressing CD19sIg1-4 (D) . Lanes E and F are the DYNAL purified extracts for CD19sIg1-3 and CD19sIg1-4 (respectively). Arrows indicate the predicted size of the monomer, 47 kDa for CD19sIg1-3 and 57 kDa for CD19sIg1-4. All samples were reduced prior to loading. C . Native protein electrophoresis of CD19sIg1-4 after concentration and purification using Protein A Dynabeads; arrow points to 171 kDa band (expected size for trimers of the fusion protein). D . Results of a comparative ELISA using FMC63 monoclonal capture antibody, of purified fusion proteins, in native (N) or denatured (D) forms. E . Results of ELISA using antibodies targeting human CD19 molecule (FMC63, HIB19, F-3 and 2E2B6B10), human CD20 (B9E9) and PSMA (YPSMA-1).

Article Snippet: The CD19-IgG 1 Fc fusion proteins, CD19sIg1-3 and CD19sIg1-4, were constructed by fusing either exons 1 to 3 (E13) or exons 1 to 4 (E14) of the human CD19 cDNA (Origene, Rockville, MD) to a human IgG 1 Fc (Fc) fragment [ ] by PCR-based cloning.

Techniques: Plasmid Preparation, Construct, Transduction, Protein Electrophoresis, SDS Page, Concentration Assay, Purification, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors

doi: 10.1186/1479-5876-11-23

Figure Lengend Snippet: Evaluation of CD19sIg1-4 fusion protein on primary human cell populations. Flow cytometry plots using FITC-conjugated anti-IgG Fc F(ab’) 2 fragment (FITC-anti-IgG Fc, left panels) or Alexa Fluor 488-labeled CD19sIg1-4 (AF488-CD19sIg1-4, right panels) for detection of 5% anti-CD19 CAR-transduced human primary T-cells mixed with (A) human peripheral blood mononuclear cells (PBMC), (B) NSG bone marrow (NSG BM), and (C) humanized NSG bone marrow ( hu NSG BM).

Article Snippet: The CD19-IgG 1 Fc fusion proteins, CD19sIg1-3 and CD19sIg1-4, were constructed by fusing either exons 1 to 3 (E13) or exons 1 to 4 (E14) of the human CD19 cDNA (Origene, Rockville, MD) to a human IgG 1 Fc (Fc) fragment [ ] by PCR-based cloning.

Techniques: Flow Cytometry, Labeling

Journal: Journal of Translational Medicine

Article Title: A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors

doi: 10.1186/1479-5876-11-23

Figure Lengend Snippet: Evaluation of CD19sIg1-4 fusion protein for detection of anti-CD19 CAR-modified primary human T-cells. Flow cytometry plots demonstrating the sensitivity of detection of anti-CD19 CAR-transduced human primary T-cells mixed in increasing numbers of non-transduced (NT) T-cells using FITC-conjugated anti-IgG Fc F(ab’) 2 fragment (FITC-anti-IgG Fc, upper panels) or Alexa Fluor 488-labeled CD19sIg1-4 (AF488-CD19sIg1-4, lower panels).

Article Snippet: The CD19-IgG 1 Fc fusion proteins, CD19sIg1-3 and CD19sIg1-4, were constructed by fusing either exons 1 to 3 (E13) or exons 1 to 4 (E14) of the human CD19 cDNA (Origene, Rockville, MD) to a human IgG 1 Fc (Fc) fragment [ ] by PCR-based cloning.

Techniques: Modification, Flow Cytometry, Labeling

Journal: Journal of Translational Medicine

Article Title: A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors

doi: 10.1186/1479-5876-11-23

Figure Lengend Snippet: Comparison of CD19sIg1-4 fusion protein to similar commercially available reagents. Flow cytometry plots of staining of primary human T-cells, non-transduced and 5% CAR-transduced, using FITC-conjugated F(ab’) 2 fragment goat anti-human IgG1 Fc γ (A) , biotinylated Protein L (B) , Alexa Fluor 488-labeled CD19sIg1-4 (C) and Alexa Fluor 488-labeled rhCD19-Fc fusion protein (D) . E . Results of staining of the same cell population after pre-incubation of Alexa Fluor 488-labeled CD19sIg1-4 with anti-CD19 monoclonal antibody FMC63.

Article Snippet: The CD19-IgG 1 Fc fusion proteins, CD19sIg1-3 and CD19sIg1-4, were constructed by fusing either exons 1 to 3 (E13) or exons 1 to 4 (E14) of the human CD19 cDNA (Origene, Rockville, MD) to a human IgG 1 Fc (Fc) fragment [ ] by PCR-based cloning.

Techniques: Flow Cytometry, Staining, Labeling, Incubation

Journal: Molecular cell

Article Title: Dynamic Processing of Displacement Loops during Recombinational DNA Repair

doi: 10.1016/j.molcel.2019.01.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-Myc (9E11) Santa Cruz Biotechnology sc-47694, lot F11 13; RRID: AB_627266 Mouse monoclonal anti-GAPDH Thermo Scientific MA5–15738, lot QG215126; RRID: AB_10977387 Chemicals, Peptides, and Recombinant Proteins Trioxsalen (TMP) Sigma-Aldrich Cat#T6137 lndole-3-acetic acid (IAA) sodium salt (Auxin) Sigma-Aldrich Cat#l5148 dCTP-alpha-P32 6000 uCi/mmol PerkinElmer Cat#BLU-513Z T4 DNA Ligase New England Biolabs Cat#M0202 EcoRI-HF New England Biolabs Cat#R3101 Hindlll-HF New England Biolabs Cat#R3104 Critical Commercial Assays LightCycler 480 SYBR Green I Master Roche Life Science Cat#04707516001 Ambion DecaPrime II DNA labeling kit Thermo Fisher Cat#AM1455 Experimental Models: Organisms/Strains Saccharomyces cerevisiae; Individual genotypes see Table S1 This study N/A Oligonucleotides Quantitative PCR primers, see Table S2 Eurofins Genomics N/A Hybridization oligonucleotides, see Table S2 Sigma-Aldrich N/A Software and Algorithms LightCycler 480 Software Roche Life Science Cat#04994884001 R ×64 3.2.0 R Project https://www.r-project.org/ Ape Plasmid Editor Wayne Davis https://biologylabs.utah.edu/jorgensen/wayned/ape/ ImageJ 1.51k Wayne Rasband https://imagej.nih.gov/ij/ Other Spectrolinker (with 365 nm UV bulbs) Spectroline XL-1500 Open in a separate window KEY RESOURCES TABLE Development of a physical assay for D-loop detection in cells Srs2, Mph1, and Sgs1-Top3-Rmi1 (STR) regulate D-loop levels in vivo Two distinct pathways (Srs2 and Mph1, STR) target different D-loop species Rdh54 delineates the two D-loop reversal pathways

Techniques: Recombinant, SYBR Green Assay, DNA Labeling, Real-time Polymerase Chain Reaction, Hybridization, Software, Plasmid Preparation