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    Hypoxia Reporter Element Assay
    Authors:  Daelynn R Buelow and Sharyn D. D Baker, date: 08/05/2018, view: 77, Q&A: 0
    Hypoxia is a condition in which there is a decrease in oxygen supply to the cellular environment. Changes to cellular oxygen levels can lead to transcriptional changes of oxygen-regulated genes. Reporter assays are used to study gene expression ...
    Hypoxia Reporter Element Assay
    Authors:  Daelynn R Buelow and Sharyn D. D Baker, date: 08/05/2018, view: 77, Q&A: 0
    [Abstract] Hypoxia is a condition in which there is a decrease in oxygen supply to the cellular environment. Changes to cellular oxygen levels can lead to transcriptional changes of oxygen-regulated genes. Reporter assays are used to study gene expression alteration and modifications in response to environmental changes. Dual-reporter assays allow the ...
    An in vitro Co-culture System for the Activation of CD40 by Membrane-presented CD40 Ligand versus Soluble Agonist
    [Abstract] One fundamental property of the TNR receptor (TNFR) family relates to how ‘signal quality’ (the extent of receptor ligation or cross-linking) influences the outcome of receptor ligation, for instance the induction of death in tumour cells. It is unequivocal that membrane-presented ligand (delivered to target cells via cell-surface presentation by ...
    Mouse Mammary Gland Whole Mount Preparation and Analysis
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 07/05/2018, view: 451, Q&A: 0
    [Abstract] The mammary gland undergoes extensive remodeling during pregnancy and is also subject to neoplastic processes both of which result in histological changes of the gland epithelial structure. Since the mammary tree is a complex three-dimensional structure a method is needed that provides an overview of the entire gland. Whole mounts provide this ...
    Isolation of Microvascular Endothelial Cells
    Authors:  Kenneth C.P. Cheung and Federica M. Marelli-Berg, date: 06/20/2018, view: 827, Q&A: 0
    [Abstract] The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. Murine endothelial cell culture is an important tool for cardiovascular disease research. This protocol demonstrates a quick, efficient method for the isolation of microvascular endothelial cells from murine ...
    3D Culture Protocol for Testing Gene Knockdown Efficiency and Cell Line Derivation
    Authors:  Jan Strnadel, Sang Myung Woo, Sunkyu Choi, Huawei Wang, Marian Grendar and Ken Fujimura, date: 06/05/2018, view: 805, Q&A: 0
    [Abstract] Traditional 2D cell cultures with cells grown as monolayers on solid surface still represent the standard method in cancer research for drug testing. Cells grown in 2D cultures, however, lack relevant cell-matrix and cell-cell interactions and ignore the true three-dimensional anatomy of solid tumors. Cells cultured in 2D can also undergo ...
    Hyaluronan Isolation from Mouse Mammary Gland
    Authors:  Cornelia Tolg, Mary Cowman and Eva A. Turley, date: 06/05/2018, view: 429, Q&A: 0
    [Abstract] The glycosaminoglycan hyaluronan (HA) is a key component of the extracellular matrix. The molecular weight of HA is heterogeneous and can reach from several million to several hundred daltons. The effect of HA on cell behavior is size dependent; fragmented HA acts as a danger signal, stimulates cell migration and proliferation and is ...
    Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism
    Authors:  Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 2182, Q&A: 0
    [Abstract] Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...
    An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells
    Authors:  Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 1017, Q&A: 0
    [Abstract] In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...
    Generation of Luciferase-expressing Tumor Cell Lines
    Authors:  Todd V. Brennan, Liwen Lin, Xiaopei Huang and Yiping Yang, date: 04/20/2018, view: 1588, Q&A: 1
    [Abstract] Murine tumor models have been critical to advances in our knowledge of tumor physiology and for the development of effective tumor therapies. Essential to these studies is the ability to both track tumor development and quantify tumor burden in vivo. For this purpose, the introduction of genes that confer tumors with bioluminescent ...
    3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells
    Authors:  Lingli Long, Mingzhu Yin and Wang Min, date: 04/20/2018, view: 1239, Q&A: 0
    [Abstract] Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated ...