deadend tm fluoremetric tunel kit #tb235  (Promega)

Journal: Nature Communications

Article Title: Pervasive structural heterogeneity rewires glioblastoma chromosomes to sustain patient-specific transcriptional programs

doi: 10.1038/s41467-024-48053-2

Figure Lengend Snippet: a Exemplary Hi-C contact map from G148 around a translocation breakpoint involving the MYC locus. Enhancer-promoter neoloops forming across the breakpoint are indicated (blue circles) and aligned to RNA-seq, H3K27ac, and CNV tracks. Absence of neoloops in G394 (below), where no translocation occurs, provides a control. b Plot showing mean (line) ±SEM GSC-specific MYC expression (circles) in cells with (orange; n = 1) or without the chr8:chr12 translocation (gray; n = 9) or in astrocytes (black; n = 2). Source data for this panel are provided as a Source Data file. c , Plots showing the percentage of cells ±SD staining positive for MYC or Ki-67 in untreated (red) or EN4-treated MYC -high G148 (green) from n = 4 independent experiments; treatment of the MYC -low G62 provides a control. *: P = 0.038 (for MYC) and 0.021 (for Ki-67), unpaired two-sided Student’s t-test. Source data for this panel are provided as a Source Data file. d As in panel ( c ) but showing the percentage of cells ±SD positive for TUNEL staining from n = 4 independent experiments. *: P = 0.00025, unpaired two-sided Student’s t-test. Source data for this panel are provided as a Source Data file. e Heatmaps showing correlation of each class of polymer beads with H3K27ac and RNA-seq data from G148 that carries the chr8:chr12 translocation (left) or G394 that does not (right). Classes with a correlation of >0.2 are shown. f Left: Contact maps from Hi-C (top) or simulations (bottom) around the translocation breakpoint in G148 are shown aligned to polymer bead classes. Enhancer-promoter neoloops forming across the breakpoint are indicated (blue circles). Right: As in the left hand-side panel, but for G394 that does not carry the translocation. g Representative 3D renderings of the two major configurations resulting from chr8:chr12 ecDNA translocation involving the MYC locus. Beads from binding classes 1, 7, and 11 that best predict folding are color-coded as in panel d, and differential MYC -enhancer interactions indicated (yellow halo).

Article Snippet: For TUNEL stainings, the DeadEnd TM Fluoremetric TUNEL kit (#TB235, Promega) was used according to the manufacturer’s instructions.

Techniques: Hi-C, Translocation Assay, RNA Sequencing, Control, Expressing, Staining, TUNEL Assay, Polymer, Binding Assay

Journal: iScience

Article Title: Heterogeneity of perivascular astrocyte endfeet depending on vascular regions in the mouse brain

doi: 10.1016/j.isci.2023.108010

Figure Lengend Snippet:

Article Snippet: DeadEnd Fluoremetric TUNEL System , Promega , Cat# G3250.

Techniques: Produced, Recombinant, Affinity Purification, Protease Inhibitor, Western Blot, Fluorsave, Bicinchoninic Acid Protein Assay, Double Staining, TUNEL Assay, Mass Spectrometry, Plasmid Preparation, Software

Journal: Frontiers in Plant Science

Article Title: PsEND1 Is a Key Player in Pea Pollen Development Through the Modulation of Redox Homeostasis

doi: 10.3389/fpls.2021.765277

Figure Lengend Snippet: Phenotypic characterization of VIGS- PsEND1 pea flowers. (A) Expression of PsEND1 in flower buds of different VIGS plants. (B) Detail of the stamens of wild type (upper) and VIGS- PsEND1 (bottom) plants. The anthers of the VIGS- PsEND1 plants showed a white color and small size. (C) Percentage of viable pollen in the wild type and VIGS- PsEND1 plants. (D) Alexander’s staining of pollen; wild type (upper) and VIGS plant (bottom). Viable pollen protoplasm is stained pink, while dead pollen is stained blue. Histological sections of wild type (E–G) and VIGS- PsEND1 (H–J) pea flowers. (E,H) 3 mm flowers, (F,I) 5 mm flowers, (G,J) 7 mm flowers. Arrows in (E–H) indicate the presence of tapetal cells. Scale bars in (E–J) correspond to 100 μm. (K–P) Detection of DNA fragmentation by TUNEL assays of pea anthers of the wild type (K–M) and VIGS- PsEND1 anthers (N–P) . Cell wall or membranes showed red fluorescence and the positive apoptotic nuclei stained with TUNEL were deep green.

Article Snippet: TdT-mediated dUTP Nick-End Labeling (TUNEL) assay was performed with the DeadEND TM Fluoremetric TUNEL System kit (Promega) according to the manufacturer instructions.

Techniques: Expressing, Staining, TUNEL Assay, Fluorescence

Journal: Molecular Therapy

Article Title: Systemic Delivery of Modified mRNA Encoding Herpes Simplex Virus 1 Thymidine Kinase for Targeted Cancer Gene Therapy

doi: 10.1038/mt.2012.250

Figure Lengend Snippet: Systemic delivery of modified mRNA encoding HSV-tk in the LPR coupled with GCV for cancer gene therapy. (a) MTS cell proliferation assay used to determine the killing efficiency of LPR (HSV-tk)/GCV in vitro. The data were reported as mean ± SD. (b) Flow cytometry analysis of H460 cells treated with LPR (HSV-tk)/GCV followed by PI/annexin V staining. Cells were transfected with LPR (HSV-tk) nanoparticles in combination with GCV. Timeline of cell apoptosis was determined by PI/annexin-V double staining followed by flow cytometry analysis. (c) Proliferation capacity for the survival cells after HSV-tk/GCV therapy determined by clonogenic assay. The number was normalized against untreated group. The data were reported as mean ± SD. (d) Tumor growth inhibition study on H460 tumor xenograft. Tumor size was monitored every 3 days during the drug treatment. Tumor volumes were calculated as ½ × length × width × height. The data were reported as mean ± SEM (n = 4–6). (e) Tumor volumes from animals at end point of the experiment. Each symbol represents an individual mouse. Horizontal lines indicate mean values. (f) TUNEL assay showing the apoptosis degree of tumors harvested from end point of the tumor growth inhibition experiment. Con, control; GCV, ganciclovir; HSV-tk, herpes simplex virus-thymidine kinase; LPR, lipid/protamine/mRNA; mRNA, messenger RNA; PI, propidium iodide; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

Article Snippet: After deparaffinization, section was stained with DeadEnd Fluoremetric TUNEL Assay kit (Promega) according to the manufacturer's manual.

Techniques: Modification, Proliferation Assay, In Vitro, Flow Cytometry, Staining, Transfection, Double Staining, Clonogenic Assay, Inhibition, TUNEL Assay, Control, Virus