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Cancer Biology

Analysis of Cancer Stromal Reaction Using an O-ring Co-culture Assay Author:  Vivien Jane Coulson-Thomas, date: 02/20/2017, view: 2755, Q&A: 0
We have developed a 2D heterotypic co-culture technique between fibroblasts and cancer cells that enables the study of the stromal reaction. For such, stromal cells are seeded and cultured immediately around a tumour cell line, and the cells establish cell-cell contacts, as well as a gradient of soluble factors throughout the stromal cells, ...
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Protocol for Murine/Mouse Platelets Isolation and Their Reintroduction in vivo Authors:  Jae Hong Im and Ruth J. Muschel, date: 02/20/2017, view: 4847, Q&A: 0
Platelets and coagulation have long been known to be essential for metastasis in experimental models. In order to study the interactions between tumor cells, platelets and endothelium, we have adapted methods used in coagulation research for the isolation of platelets and their reintroduction into mice. Anti-coagulated murine blood served as the ...
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A Murine Orthotopic Allograft to Model Prostate Cancer Growth and Metastasis Authors:  Robert M. Hughes, Brian W. Simons and Paula J. Hurley, date: 02/20/2017, view: 3483, Q&A: 0
Prostate cancer is one of the most common cancers in men in the United States. Comprehensive understanding of the biology contributing to prostate cancer will have important clinical implications. Animal models have greatly impacted our knowledge of disease and will continue to be a valuable resource for future studies. Herein, we describe a ...
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In situ Hybridization (ISH) and Quantum Dots (QD) of miRNAs Authors:  Sajni Josson, Murali Gururajan and Leland W.K. Chung, date: 02/20/2017, view: 3217, Q&A: 0
miRNA are short non-coding RNA which inhibit translation of mRNA. miRNA regulate several cellular processes. Certain miRNA are known to induce oncogenesis. miRNA can be measured by real-time PCR and be imaged using a combination of in situ hybridization (ISH) and quantum dots (QD). The advantage of using quantum dots is that several miRNA ...
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miRNA Characterization from the Extracellular Vesicles Authors:  Sajni Josson, Murali Gururajan and Leland W.K. Chung, date: 02/20/2017, view: 2310, Q&A: 0
Cancer cells and cancer associated stromal cells co-evolve secrete extracellular vesicles to the surrounding regions and regulate several processes involved in cancer metastasis. miRNAs have been known to be mediators of cancer progression and metastasis. miRNAs consist of short noncoding RNA. miRNAs are stable in extracellular fluids such as ...
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Cell Biology

Quantitative Analysis of Exosome Secretion Rates of Single Cells Authors:  Yu-Jui Chiu, Wei Cai, Tiffany Lee, Julia Kraimer and Yu-Hwa Lo, date: 02/20/2017, view: 3934, Q&A: 0
To study the inhomogeneity within a cell population including exosomes properties such as exosome secretion rate of cells and surface markers carried by exosomes, we need to quantify and characterize those exosomes secreted by each individual cell. Here we develop a method to collect and analyze exosomes secreted by an array of single cells using ...
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Microbiology

Expression and Purification of Cyanobacterial Circadian Clock Protein KaiC and Determination of Its Auto-phosphatase Activity Authors:  Qiang Chen, Lingling Yu, Xiao Tan and Sen Liu, date: 02/20/2017, view: 2512, Q&A: 0
Circadian rhythms are biological processes displaying an endogenous oscillation with a period of ~24 h. They allow organisms to anticipate and get prepared for the environmental changes caused mainly by the rotation of Earth. Circadian rhythms are driven by circadian clocks that consist of proteins, DNA, and/or RNA. Circadian clocks of ...
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Molecular Biology

Production of Guide RNAs in vitro and in vivo for CRISPR Using Ribozymes and RNA Polymerase II Promoters Authors:  Tao Zhang, Yangbin Gao, Rongchen Wang and Yunde Zhao, date: 02/20/2017, view: 4941, Q&A: 0
CRISPR/Cas9-mediated genome editing relies on a guide RNA (gRNA) molecule to generate sequence-specific DNA cleavage, which is a prerequisite for gene editing. Here we establish a method that enables production of gRNAs from any promoters, in any organisms, and in vitro (Gao and Zhao, 2014). This method also makes it feasible to conduct ...
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Plant Science

Penetration Assays, Fungal Recovery and Pathogenicity Assays for Verticillium dahliae Authors:  Yun-Long Zhao, Tao Zhang and Hui-Shan Guo, date: 02/20/2017, view: 3130, Q&A: 0
Verticillium dahliae is a soil-borne phytopathogenic fungus that infects host roots and proliferates in vascular tissues. The great loss of economically important crop caused by V. dahliae has raised worldwide concern, however, little is known about the mechanism of its pathogenicity (Klosterman et al., 2011; Yadeta and ...
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Establishment of New Split-root System by Grafting Authors:  Xiangqiang Kong, Zhen Luo and Hezhong Dong, date: 02/20/2017, view: 2590, Q&A: 0
A new split-root system was used to simulate non-uniform salt, drought or nutrient deficiency stress in the root zone, in which the root system was divided into two or more equal portions. Here, we established a split-root system by grafting of cotton seedlings. In contrast to the conventional split-root, the main roots of the new system remained ...
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Rice Root Organic Acid Efflux Measurement by Using Ion Chromatography Authors:  Chun-quan Zhu, Xiao-fang Zhu and Ren-fang Shen, date: 02/20/2017, view: 2601, Q&A: 0
Organic acids secreted from plant roots play important roles in various biological processes including nutrient acquisition, metal detoxification, and pathogen attraction. The secretion of organic acids may be affected by various conditions such as plant growth stage, nutrient deficiency, and abiotic stress. For example, when white lupin (Lupinus ...
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Endophytic Microbial Community DNA Extraction from the Plant Phyllosphere Authors:  Carlos A. Ruiz-Pérez and María Mercedes Zambrano, date: 02/20/2017, view: 2947, Q&A: 1
The plant phyllosphere, which represents all plant parts that are above the ground, is considered one of the most extensive ecosystems to be colonized by microorganisms, both at the surface as epiphytes or as endophytes within the plant. These plant-associated microbial communities are reservoirs of microbial diversity and they can be important ...
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Chromatographic Separation of the Codonocarpine Type Alkaloids from the Root Bark of Capparis decidua Authors:  Yvonne Forster, Abdul Ghaffar and Stefan Bienz, date: 02/20/2017, view: 2465, Q&A: 0
Various parts of the caper tree Capparis decidua have found application in traditional medicine. The isolation and structural elucidation of the codonocarpine type alkaloids contained in the root bark, however, is not trivial and has probably led to misinterpretation in the past. This protocol describes the extraction and chromatographic ...
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Automatic Quantification of the Number of Intracellular Compartments in Arabidopsis thaliana Root Cells Authors:  Vincent Bayle, Matthieu Pierre Platre and Yvon Jaillais, date: 02/20/2017, view: 3724, Q&A: 0
In the era of quantitative biology, it is increasingly required to quantify confocal microscopy images. If possible, quantification should be performed in an automatic way, in order to avoid bias from the experimenter, to allow the quantification of a large number of samples, and to increase reproducibility between laboratories. In this protocol, ...
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Stem Cell

VLA-4 Affinity Assay for Murine Bone Marrow-derived Hematopoietic Stem Cells Authors:  Francesca Avemaria, Shiri Gur-Cohen, Seymen Avci and Tsvee Lapidot, date: 02/20/2017, view: 2506, Q&A: 0
Hematopoietic stem cells (HSCs) are defined by their functional ability to self-renew and to differentiate into all blood cell lineages. The majority of HSC reside in specific anatomical locations in the bone marrow (BM) microenvironment, in a quiescent non motile mode. Adhesion interactions between HSCs and their supporting BM microenvironment ...
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Adhesion Assay for Murine Bone Marrow Hematopoietic Stem Cells Authors:  Seymen Avci, Shiri Gur-Cohen, Francesca Avemaria and Tsvee Lapidot, date: 02/20/2017, view: 3159, Q&A: 0
Hematopoietic stem cells (HSCs) are defined by their functional abilities to self-renew and to give rise to all mature blood and immune cell types throughout life. Most HSCs are retained in a non-motile quiescent state within a specialized protective microenvironment in the bone marrow (BM) termed the niche. HSCs are typically distinguished from ...
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