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Mapping the Mechanome–A Protocol for Simultaneous Live Imaging and Quantitative Analysis of Cell Mechanoadaptation and Ingression

Authors: Vina D. L. Putra, Iman Jalilian, Madeline Campbell, Kate Poole, Renee Whan, Florence Tomasetig and Melissa L. Knothe Tate, date: 12/05/2019, view: 87, Q&A: 0

Mechanomics, the mechanics equivalent of genomics, is a burgeoning field studying mechanical modulation of stem cell behavior and lineage commitment. Analogous to mechanical testing of a living material as it adapts and evolves, mapping of the ...

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Mapping the Mechanome–A Protocol for Simultaneous Live Imaging and Quantitative Analysis of Cell Mechanoadaptation and Ingression

Authors: Vina D. L. Putra, Iman Jalilian, Madeline Campbell, Kate Poole, Renee Whan, Florence Tomasetig and Melissa L. Knothe Tate, date: 12/05/2019, view: 87, Q&A: 0

[Abstract] Mechanomics, the mechanics equivalent of genomics, is a burgeoning field studying mechanical modulation of stem cell behavior and lineage commitment. Analogous to mechanical testing of a living material as it adapts and evolves, mapping of the mechanome necessitates the development of new protocols to assess changes in structure and function in ...

Isolation and Quantification of Metabolite Levels in Murine Tumor Interstitial Fluid by LC/MS

Authors: Mark R Sullivan, Caroline A Lewis and Alexander Muir, date: 11/20/2019, view: 668, Q&A: 0

[Abstract] Cancer is a disease characterized by altered metabolism, and there has been renewed interest in understanding the metabolism of tumors. Even though nutrient availability is a critical determinant of tumor metabolism, there has been little systematic study of the nutrients directly available to cancer cells in the tumor microenvironment. Previous ...

Probabilistic Models for Predicting Mutational Routes to New Adaptive Phenotypes

Authors: Eric Libby and Peter A. Lind, date: 10/20/2019, view: 528, Q&A: 0

[Abstract] Understanding the translation of genetic variation to phenotypic variation is a fundamental problem in genetics and evolutionary biology. The introduction of new genetic variation through mutation can lead to new adaptive phenotypes, but the complexity of the genotype-to-phenotype map makes it challenging to predict the phenotypic effects of ...

Non-aqueous Fractionation (NAF) for Metabolite Analysis in Subcellular Compartments of Arabidopsis Leaf Tissues

Authors: David B. Medeiros, Stéphanie Arrivault, Jessica Alpers, Alisdair R. Fernie and Fayezeh Arabi, date: 10/20/2019, view: 367, Q&A: 0

[Abstract] The accurate determination of metabolite distribution in subcellular compartments is still challenging in plant science. Various methodologies, such as fluorescence resonance energy transfer-based technology, nuclear magnetic resonance spectroscopy and protoplast fractionation allow the study of metabolite compartmentation. However, large changes ...

Electron Microscopy Sample Preparation Protocol Enabling Nano-to-mesoscopic Mapping of Cellular Connectomes and Their Habitats in Human Tissues and Organs

Authors: Lucy Ngo, Anton D. Nathanson, Tomasz Garbowski, Ulf Knothe, Dirk Zeidler and Melissa L. Knothe Tate, date: 07/20/2019, view: 1066, Q&A: 0

[Abstract] Multibeam scanning electron microscopy (multiSEM) provides a technical platform for seamless nano-to-mesoscale mapping of cells in human tissues and organs, which is a major new initiative of the U.S. National Institutes of Health. Such cross-length-scale imaging is expected to provide unprecedented understanding of relationships between cellular ...

A Protocol to Map the Spatial Proteome Using HyperLOPIT in Saccharomyces cerevisiae

Authors: Daniel J. H. Nightingale, Kathryn S. Lilley and Stephen G. Oliver, date: 07/20/2019, view: 1010, Q&A: 0

[Abstract] The correct subcellular localization of proteins is vital for cellular function and the study of this process at the systems level will therefore enrich our understanding of the roles of proteins within the cell. Multiple methods are available for the study of protein subcellular localization, including fluorescence microscopy, organelle ...

Genome-wide Estimation of Evolutionary Distance and Phylogenetic Analysis of Homologous Genes

Authors: Meixia Zhao, Biao Zhang, Jianxin Ma and Damon Lisch, date: 12/05/2018, view: 2265, Q&A: 0

[Abstract] Homologous genes, including paralogs and orthologs, are genes that share sequence homologies within or between different species. Homologous genes originate from a common origin through speciation, genetic duplication or horizontal gene transfer. Estimation of the sequence divergence of homologous genes help us to understand divergence time, which ...

Use of Gas Chromatography to Quantify Short Chain Fatty Acids in the Serum, Colonic Luminal Content and Feces of Mice

Authors: Willian Rodrigues Ribeiro, Marco Aurélio Ramirez Vinolo, Leandro Augusto Calixto and Caroline Marcantonio Ferreira, date: 11/20/2018, view: 4659, Q&A: 0

[Abstract] Short-Chain Fatty Acids (SCFAs) are a product of the fermentation of resistant starches and dietary fibers by the gut microbiota. The most important SCFA are acetate (C2), propionate (C3) and butyrate (C4). These metabolites are formed and absorbed in the colon and then transported through the hepatic vein to the liver. SCFAs are more concentrated ...

Stable-isotope Labeled Metabolic Analysis in Drosophila melanogaster: From Experimental Setup to Data Analysis

Authors: Yuping Cai, Nan Liu and Zheng-Jiang Zhu, date: 09/20/2018, view: 2268, Q&A: 0

[Abstract] Stable-isotope labeled metabolic analysis is an essential methodology to characterize metabolic regulation during biological processes. However, the method using stable-isotope-labeled tracer (e.g., ¹³C-glucose) in live animal is only beginning to be developed. Here, we contribute a qualitative metabolic labeling experiment ...

Quantitative ChIP-seq by Adding Spike-in from Another Species

Authors: Kongyan Niu, Rui Liu and Nan Liu, date: 08/20/2018, view: 4414, Q&A: 0

[Abstract] Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a routine procedure in the lab; however, epigenome-wide quantitative comparison among independent ChIP-seq experiments remains a challenge. Here, we contribute an experimental protocol combined with a computational workflow allowing quantitative and comparative assessment of ...

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