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ATAC-seq on Sorted Adult Mouse Neurons

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SIRF: A Single-cell Assay for in situ Protein Interaction with Nascent DNA Replication Forks

Authors: Sunetra Roy and Katharina Schlacher, date: 09/20/2019, view: 308, Q&A: 0

[Abstract] The duplication of DNA is a fundamental process that is required for the transfer of the genetic information from parent to daughter cells. Aberrant DNA replication processes are associated with diverse disease phenotypes, including developmental defects, ageing disorders, blood disorders such as Fanconi Anemia, increased inflammation and cancer. ...

Optimized Protocol for the Incorporation of FDAA (HADA Labeling) for in situ Labeling of Peptidoglycan

Authors: Katharina Peters, Manuel Pazos, Michael S. VanNieuwenhze and Waldemar Vollmer, date: 08/05/2019, view: 535, Q&A: 0

[Abstract] The essential peptidoglycan (PG) layer surrounds the cytoplasmic membrane in nearly all bacteria. It is needed to maintain the shape of the cell and protect it from lysis due to high turgor. Growth of the PG layer is a complex process that involves the activities of PG synthases and hydrolases during elongation and cell division. PG growth sites ...

Endocytosis Detection in Magnaporthe oryzae

Authors: Muxing Liu and Zhengguang Zhang, date: 08/05/2019, view: 398, Q&A: 0

[Abstract] Endocytosis is an intracellular trafficking pathway that occurs in nutrient uptake, signal transduction and reconstruction of cell polarity and is conserved in eukaryotic cells. In fungi, endocytosis plays crucial roles in the physiology of hyphal growth and pathogenicity. vidence for endocytosis in filamentous fungi is detected by the ...

Evaluation of Genotoxicity by Micronucleus Assay in vitro and by Allium cepa Test in vivo

Authors: Christina N. Banti and Sotiris K. Hadjikakou, date: 07/20/2019, view: 955, Q&A: 0

[Abstract] The in vitro and in vivo genotoxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) is evaluated by the micronucleus test and the Allium cepa assay, respectively. Fetal lung fibroblast cells (MRC-5), normal human corneal epithelial cells (HCEC) and ...

Plaque Assay to Determine Invasion and Intercellular Dissemination of Shigella flexneri in TC7 Human Intestinal Epithelial Cells

Authors: Atin Sharma and Andrea Puhar, date: 07/05/2019, view: 626, Q&A: 0

[Abstract] Shigella flexneri invades the epithelial cells lining the gut lumen and replicates intracellularly. The specialized Type III Secretion System (T3SS) and its effector proteins, encoded on a large virulence plasmid, assist the bacterium to gain access to the cytosol. Thereafter Shigella disseminates to neighboring cells in an ...

Visualizing Filamentous Actin in Chlamydomonas reinhardtii

Authors: Evan W. Craig and Prachee Avasthi, date: 06/20/2019, view: 1030, Q&A: 0

[Abstract] This protocol aims to visualize the filamentous actin network in Chlamydomonas reinhardtii. We improved fixed-cell labeling conditions using the F-actin probe, phalloidin. We created a Chlamydomonas-optimized protocol by halving the phalloidin incubation time, electing for optimal fixation conditions, and selecting for a healthy ...

Protocol to Quantitatively Assess the Structural Integrity of Perineuronal Nets ex vivo

Authors: Bhanu P. Tewari and Harald Sontheimer, date: 05/20/2019, view: 1079, Q&A: 0

[Abstract] Perineuronal nets (PNNs) are extracellular matrix assemblies of highly negatively charged proteoglycans that wrap around fast-spiking parvalbumin (PV) expressing interneurons in the cerebral cortex. PNNs play important roles in neuronal plasticity and modulate biophysical properties of the enclosed interneurons. Various central nervous system ...

Measuring Protein Synthesis during Cell Cycle by Azidohomoalanine (AHA) Labeling and Flow Cytometric Analysis

Authors: Koshi Imami and Tomoharu Yasuda, date: 04/20/2019, view: 1483, Q&A: 0

[Abstract] Protein synthesis is one of the most fundamental biological processes to maintain cellular proteostasis. Azidohomoalaine (AHA) is a non-radioactive and “clickable” amino acid analog of methionine which can be incorporated into newly synthesized proteins. Thus, AHA-labeled nascent proteins can be detected and quantified through fluorescent labeling ...

High-resolution Immunoelectron Microscopy Techniques for Revealing Distinct Subcellular Type 1 Cannabinoid Receptor Domains in Brain

Authors: Nagore Puente, Itziar Bonilla-Del Río, Svein Achicallende, Patrick C. Nahirney and Pedro Grandes, date: 01/20/2019, view: 1832, Q&A: 0

[Abstract] Activation of type 1 cannabinoid (CB₁) receptors by endogenous, exogenous (cannabis derivatives) or synthetic cannabinoids (i.e., CP 55.940, Win-2) has a wide variety of behavioral effects due to the presence of CB₁ receptors in the brain. In situ hybridization and immunohistochemical techniques have been ...

Flow Cytometry Assay for Recycling of LFA-1 in T-lymphocytes

Authors: Katarzyna Potrzebowska, Janne Lehtonen, Malin Samuelsson and Lena Svensson, date: 12/05/2018, view: 1365, Q&A: 0

[Abstract] To enable cells to move forward, cell surface integrins are internalized into an endosomal compartment and subsequently intracellularly transported to be re-exposed at a new site on the cell membrane. Leukocytes are the fastest migrating cell type in the human body, which express the leukocyte-specific integrin LFA-1. Here, we describe a flow ...

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