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4 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications
Jason M. Conley 1, Tarsis F. Brust 1, Ruqiang Xu 1, Kevin D. Burris 2, Val J. Watts 1
1Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 2Quantitative Biology, Eli Lilly and Company

Persistent activation of inhibitory G protein-coupled receptors results in sensitization of adenylyl cyclase signaling. To identify the essential molecular pathways, nonbiased approaches are necessary; however, this strategy requires the development of a scalable cell-based cAMP sensitization assay. Herein, we describe a sensitization assay for small molecule and siRNA screening.

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Biology

Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae
Bryce W. Buchanan 1, Michael E. Lloyd 1,2, Sarah M. Engle 1, Eric M. Rubenstein 1
1Department of Biology, Ball State University, 2Bioproduct Research & Development, Eli Lilly and Company

Protein abundance reflects the rates of both protein synthesis and protein degradation. This article describes the use of cycloheximide chase followed by western blotting to analyze protein degradation in the model unicellular eukaryote, Saccharomyces cerevisiae (budding yeast).

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JoVE Core

Open-Source Real-Time Closed-Loop Electrical Threshold Tracking for Translational Pain Research
Aidan P. Nickerson 1,2, Graeme W. T. Newton 1, James H. O'Sullivan 3, Manuel Martinez-Perez 4, Anna C. Sales 1, Gethin Williams 5, Anthony E. Pickering 1, James P. Dunham 1
1Anaesthesia, Pain, and Critical Care Sciences, School of Physiology, Pharmacology, & Neuroscience, University of Bristol, 2Eli Lilly and Company, 3Department of Computer Science, University of Bristol, 4Department of Aerospace Engineering, University of Bristol, 5Research Computing, University of Bristol

APTrack is a software plugin developed for the Open Ephys platform that enables real-time data visualization and the closed-loop electrical threshold tracking of neuronal action potentials. We have successfully used this in microneurography for human C-fiber nociceptors and mouse C-fiber and Aδ-fiber nociceptors.

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Mass Spectrometry-Based Approaches for the Analysis of Protein Higher Order Structure
Xiaoran Roger Liu 1, Yining Huang 2
1Biologics Development, Bristol Myers Squibb, 2Eli Lilly and Company

Mass Spectrometry-Based Approaches for the Analysis of Protein Higher Order Structure

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