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Tracking the Subcellular Localization of Surface Proteins in Staphylococcus aureus by Immunofluorescence Microscopy

Authors: Salvatore J. Scaffidi, Mac A. Shebes and Wenqi Yu, date: 05/20/2021, view: 1317, Q&A: 0

[Abstract]

Surface proteins of Staphylococcus aureus and other Gram-positive bacteria play essential roles in bacterial colonization and host-microbe interactions. Surface protein precursors containing a YSIRK/GXXS signal peptide are translocated across the septal membrane at mid-cell, anchored to the cell wall peptidoglycan at the cross-wall compartment,

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Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila

Authors: Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 1162, Q&A: 0

[Abstract]

Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to knock down a gene for assessing its function. Similar perturbations in gene expression can be

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Acidified Blue Ink-staining Procedure for the Observation of Fungal Structures Inside Roots of Two Disparate Plant Lineages

Authors: Jill Kowal, Elena Arrigoni and Sophie Lane, date: 10/20/2020, view: 1187, Q&A: 0

[Abstract] Identifying microscopic mycorrhizal fungal structures in roots, i.e., hyphae, vesicles and arbuscules, requires root staining procedures that are often time consuming and involves chemicals known to present health risks from exposure. By modifying established protocols, our root staining method stains roots using a safe ink- and ...

Double Labeling of PDGFR-β and α-SMA in Swine Models of Acute Kidney Injury to Detect Pericyte-to-Myofibroblast Transdifferentation as Early Marker of Fibrosis

Authors: Alessandra Stasi, Rossana Franzin, Chiara Divella, Loreto Gesualdo, Giovanni Stallone and Giuseppe Castellano, date: 10/05/2020, view: 974, Q&A: 0

[Abstract] Growing evidences suggest that peritubular capillaries pericytes are the main source of scar-forming myofibroblasts during chronic kidney disease (CKD), as well as early phases of acute kidney injury (AKI). In a swine model of sepsis and I/R (Ischemia Reperfusion) injury-induced AKI we demonstrated that renal pericytes are able to ...

Immunofluorescent Staining of Claudin-2 in Cultured Kidney Tubular Cells

Authors: Shaista Anwer and Katalin Szaszi, date: 07/20/2020, view: 1390, Q&A: 0

[Abstract] Members of the claudin family of tight junction proteins regulate paracellular permeability and modulate cell signaling. During junction remodeling, these proteins are selectively inserted into or retrieved from the tight junctions, but the control and coordination of these processes remain incompletely understood. Visualization of claudins allows ...

Microscopy-based Methods for Rosetting Assay in Malaria Research

Authors: Wenn-Chyau Lee and Laurent Rénia, date: 07/05/2020, view: 1806, Q&A: 0

[Abstract] In malaria, rosetting phenomenon is a condition where a Plasmodium-infected erythrocyte stably adheres to at least an uninfected erythrocyte. This phenomenon that occurs in all species of human malaria parasite is likely to be an immune escape mechanism for the parasite. However, it has been associated with malaria pathogenesis, possibly ...

Testing Bone Formation Induction by Calvarial Injection Assay in vivo

Authors: Narelle E McGregor, Ingrid J Poulton, Emma C Walker and Natalie A Sims, date: 03/20/2020, view: 2216, Q&A: 0

[Abstract] Bone formation occurs during embryogenesis, skeletal growth and during the process of skeletal renewal throughout life. In the process of bone formation, osteoblasts lay down a collagen-containing matrix, termed osteoid, which is gradually hardened by incorporation of mineral crystals. Although osteoblasts can be induced to differentiate and to ...

Manganese Superoxide Dismutase Activity Assay in the Yeast Saccharomyces cerevisiae

Authors: Louise Thines and Pierre Morsomme, date: 03/05/2020, view: 1720, Q&A: 0

[Abstract] Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O₂^-) into molecular oxygen (O₂) and hydrogen peroxide (H₂O₂). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature ...

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: 3400, 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: 4646, 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 ...

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