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Past Issues in 2014

Volume: 4 Issue: 13

Immunology

Phagolysosomal Trafficking Assay Author: Alka Mehra, date: 07/05/2014, view: 12729, Q&A: 0

Phagolysosomal trafficking is an important innate defense pathway that clears microbes by delivering them to lysosomes, the degradative compartment of the cell. Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, subverts this host defense mechanism by arresting maturation of the phagosome. The ability of Mtb to arrest ...

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Pulse Chase of Suspension Cells Authors: Lai-Yee Wong, QiMing Liang, Kevin Brulois and Jae Jung, date: 07/05/2014, view: 10399, Q&A: 0

Pulse-chase method is a powerful technique used to follow the dynamics of proteins over a period of time. The expression level, processing, transport, secretion or half-life of proteins can be tracked by metabolically labeling the cells, such as with radiolabeled amino acids (pulse step). This protocol describes the condition used to study the ...

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Murine in vitro Memory T Cell Differentiation Authors: Myoungjoo V. Kim, Weiming Ouyang, Will Liao, Michael Q. Zhang and Ming O. Li, date: 07/05/2014, view: 11364, Q&A: 0

Upon pathogen encounter, naïve CD8⁺ T cells are primed and undergo massive clonal expansion. A fraction of effector CD8⁺ T cells remains during the contraction phase and differentiate into memory T cells critical for mounting robust recall responses in response to secondary infection. Low frequency of memory T cells in vivo ...

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Murine in vivo CD8⁺ T Cell Killing Assay Authors: Myoungjoo V. Kim, Weiming Ouyang, Will Liao, Michael Q. Zhang and Ming O. Li, date: 07/05/2014, view: 15554, Q&A: 0

Antigen-specific killing ability of effector CD8⁺ T cells is critical for protective immunity against infection. Here, we describe in vivo cytotoxic T cell assay to examine effector function of antigen-specific CD8⁺ T cells. Mice infected with Listeria monocytogenes (L. monocytogenes) expressing ...

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Microbiology

Determination of Rifampicin-resistance Mutation Frequency and Analysis of Mutation Spectra in Mycobacteria Authors: Liang-Dong Lyu and Guo-Ping Zhao, date: 07/05/2014, view: 9719, Q&A: 0

Understanding the genetic safeguarding mechanism of Mycobacterium tuberculosis (Mtb) may help us to explain i), how Mtb survive the genetic assaults elicited by both reactive oxygen species (ROS) and reactive nitrogen species (RNS) produced by host macrophages and ii), why some strains of Mtb, e.g., Mtb strains from East Asian ...

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Neuroscience

Open-book Preparations from Chick Embryos and DiI Labeling of Commissural Axons Authors: Nicole H. Wilson and Esther T. Stoeckli, date: 07/05/2014, view: 10499, Q&A: 0

Successful neural circuit formation relies on the accurate navigation of axons towards their targets during development. Axons are guided by a combination of short-range and long-range, attractive and repulsive cues. The commissural axons of the developing spinal cord have provided an informative in vivo model for the identification of ...

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Plant Science

Grafting Arabidopsis Authors: Tonni Grube Andersen, Dacheng Liang, Barbara Ann Halkier and Rosemary White, date: 07/05/2014, view: 13540, Q&A: 0

In Arabidopsis thaliana, hypocotyl micrografting has been used to investigate transport of flowering signals, mobile silencing signals and other peptides, proteins and secondary compounds. The effects of transported signals on target tissues require that a good vascular connection is re-established across the graft junction between the ...

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Analyses of Plant Leaf Cell Size, Density and Number, as Well as Trichome Number Using Cell Counter Plugin Authors: Yan Cheng, Ling Cao, Sheng Wang, Yongpeng Li, Hong Wang and Yongming Zhou, date: 07/05/2014, view: 21885, Q&A: 2

An Arabidopsis leaf blade is composed of many layers that are sandwiched between two layers of tough skin cells (called the epidermis). Four layers (adaxial epidermis, palisade layer, spongy mesophyll and abaxial epidermis) contain specialized cells. Here we describe a quick and simple method for analyzing the size, number and density of ...

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Plant Sequence Capture Optimised for Illumina Sequencing Authors: Axel Himmelbach, Manuela Knauft and Nils Stein, date: 07/05/2014, view: 12716, Q&A: 0

Plant Sequence Capture is used for targeted resequencing of whole exomes (all exons of a genome) of complex genomes e.g. barley and its relatives (Mascher et al., 2013). Sequencing and computing costs are significantly reduced since only the greatly enriched and gene-coding part of the barley genome is targeted, that corresponds ...

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Cytokinin Analysis: Sample Preparation and Quantification Authors: Martin Schäfer, Michael Reichelt, Ian T. Baldwin and Stefan Meldau, date: 07/05/2014, view: 12705, Q&A: 0

Cytokinins are a group of phytohormones discovered about half a decade ago by Miller et al. (1955) and Skoog et al. (1965). Since then they were found to participate in many plant physiological processes, including the regulation of the source/sink transitions, plant growth and organ development, responses to environmental ...

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Extraction of Nonstructural Carbon and Cellulose from Wood for Radiocarbon Analysis Authors: Claudia I. Czimczik, Susan E. Trumbore, Xiaomei Xu, Mariah S. Carbone and Andrew D. Richardson, date: 07/05/2014, view: 7293, Q&A: 0

This method aims at isolating nonstructural organic carbon (NSC) pools, i.e. soluble sugars and starch, from wood for radiocarbon (¹⁴C) analysis at natural abundance levels (≤1 ppt).

Pools are operationally defined to 1) physically isolate pools - prohibiting the use of destructive methods, such as compound-specific ...

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Seed Coat Permeability Test: Tetrazolium Penetration Assay Authors: Sollapura J. Vishwanath, Frédéric Domergue and Owen Rowland, date: 07/05/2014, view: 11368, Q&A: 1

Seed coat permeability is important to study as it plays significant roles in seed dormancy, germination, and protection from pathogens. Here we describe a commonly used seed coat permeability test known as the tetrazolium penetration assay with a method to quantify the levels of permeability. Tetrazolium red is a cationic dye that is widely used ...

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Extraction of Ions from Leaf Sections Authors: Yuko Kurita, Satomi Kanno, Miwa Ohnishi and Tetsuro Mimura, date: 07/05/2014, view: 5817, Q&A: 0

The concentration of ions in plant cells and tissues is an important factor to determine their functions and conditions. Here, we describe the method to extract ions from leaf sections for measurements with an ion chromatogram. This method is available for not only barley but also other plant species.

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Autoradiography of Pi Distribution in Barley Seedlings Authors: Satomi Kanno, Yuko Kurita, Miwa Ohnishi and Tetsuro Mimura, date: 07/05/2014, view: 7262, Q&A: 0

Phosphorus-32 and Phosphorus-33 are radioisotopes of phosphorus. These isotopes are used to trace ionic phosphorus and phosphorus compounds. This protocol is used to follow the movement of inorganic phosphate (PO₄^3-) from a leaf tip to the rest of the plant.

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