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7 ARTICLES PUBLISHED IN JoVE

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Biology

Electron Cryotomography of Bacterial Cells
Songye Chen 1, Alasdair McDowall 1,2, Megan J. Dobro 1, Ariane Briegel 1,2, Mark Ladinsky 1,2, Jian Shi 2, Elitza I. Tocheva 1, Morgan Beeby 1,2, Martin Pilhofer 1,2, H. Jane Ding 1, Zhuo Li 1,2, Lu Gan 1, Dylan M. Morris 1, Grant J. Jensen 1,2
1Division of Biology, California Institute of Technology - Caltech, 2Howard Hughes Medical Institute, California Institute of Technology - Caltech

We illustrate here how to use electron cryotomography (ECT) to study the ultrastructure of bacterial cells in near-native states, to "macromolecular" (~4 nm) resolution.

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Biology

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting
Anton Posch 1, Jonathan Kohn 1, Kenneth Oh 1, Matt Hammond 1, Ning Liu 1
1Bio-Rad Laboratories

V3 workflow is a western blot procedure using stain-free gels. The stain-free technology allows researchers to visualize protein separation quality, to verify the transfer efficiency, and most importantly, to validate the change in the protein of interest using total protein quantification as a reliable loading control.

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Genetics

In Vitro Transcription Assays and Their Application in Drug Discovery
Xiao Yang 1, Cong Ma 1,2
1School of Environmental and Life Sciences, University of Newcastle, 2Department of Applied Biology and Chemical Technology, The State Key Laboratory of Chirosciences, The Hong Kong Polytechnic University

In this manuscript, we describe a protocol to functionally examine transcription and the inhibitory activity of antibacterial agents targeting bacterial transcription.

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Cancer Research

Monitoring Hippo Signaling Pathway Activity Using a Luciferase-based Large Tumor Suppressor (LATS) Biosensor
Taha Azad *1, Kazem Nouri *1, Helena J. Janse van Rensburg 1, Yawei Hao 1, Xiaolong Yang 1
1Department of Pathology and Molecular Medicine, Queen's University

Here we present a luciferase-based biosensor to quantify the kinase activity of large tumor suppressor (LATS)-a central kinase in the Hippo signaling pathway. This biosensor has diverse applications in basic and translational research aimed at investigating Hippo pathway regulators in vitro and in vivo.

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Neuroscience

Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5
Shuiqing Chi 1, Mijing Fang 1, Kang Li 1, Li Yang 2, Shao-tao Tang 1
1Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center

This protocol describes the process of immunostaining rectal suction biopsies for calretinin, S100 protein, and protein gene product 9.5. This novel adjuvant diagnostic method for Hirschsprung's disease has preferable sensitivity and specificity rates.

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Environment

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants
Zhuo Li 1,2, Fangting Ai 3, Jing Zhang 4, Zhenyang Yu 1,2, Daqiang Yin 1,2
1College of Environmental Sciences and Engineering, Tongji University, 2Shanghai Institute of Pollution Control and Ecological Security, 3Jiaxing Tongji Institute of Environment, 4College of Ecological Technique and Engineering, Shanghai Institute of Technology

Trans- and multi-generational effects of persistent chemicals are essential in judging their long-term consequences in the environment and on the human health. We provide novel detailed methods for studying trans- and multi-generational effects using free-living nematode Caenorhabditis elegans.

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Cancer Research

A Human Peripheral Blood Mononuclear Cell (PBMC) Engrafted Humanized Xenograft Model for Translational Immuno-oncology (I-O) Research
Zhuo Li *1, Xiao Yang *1, Yilu Zhang 1, Xiaolong Yang 1, Xinxin Cui 1, Yanjuan Zhang 1, Wenfeng Gong 1, Huichen Bai 1, Ning Liu 1, Zhiyu Tang 1, Mingming Guo 1, Kang Li 1, Tong Zhang 1, Lai Wang 1, Xiaomin Song 1
1BeiGene (Beijing) Co., Ltd.

We describe a human peripheral blood mononuclear cell (PBMC) — based humanized xenograft mouse model for translational immuno-oncology research. This protocol could serve as a general guideline for establishing and characterizing similar models for I-O therapy assessment.

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