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  1. No Access

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    Analysis of S1P Receptor Expression by Uterine Immune Cells Using Standardized Multi-parametric Flow Cytometry

    Flow cytometry is a powerful tool for phenotypic and functional analyses of single immune cells. The increasing capability of flow cytometry technology has driven a more detailed understanding of immune cell s...

    Jianhong Zhang, Annie Bang, Stephen J. Lye (2017)

  2. No Access

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    Antibody Uptake Assay in the Embryonic Zebrafish Forebrain to Study Notch Signaling Dynamics in Neural Progenitor Cells In Vivo

    Stem cells can generate cell fate heterogeneity through asymmetric cell division (ACD). ACD derives from the asymmetric segregation of fate-determining molecules and/or organelles in the dividing cell. Radial ...

    Kai Tong, Mahendra Wagle, Su Guo (2017)

  3. No Access

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    MicroRNA Expression Profiling by PCR Array in 2D and 3D Differentiated Neural Culture Systems and Target Validation

    MicroRNAs (miRNAs) have been proven to regulate gene expression at post-transcriptional level and are emerging as strong mediators in neural fate determination (Ambros, Nature 431(7006):350–355, 2004). Here, w...

    Lara Stevanato, Caroline Hicks, Lavaniya Thanabalasundaram, John D. Sinden (2017)

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    Monitoring of Paternal Mitochondrial Degradation in Caenorhabditis elegans

    In Caenorhabditis elegans embryos, paternally inherited mitochondria and their mitochondrial DNA are degraded via selective autophagy called allophagy (allogeneic organelle autophagy). This is a developmentally p...

    Miyuki Sato, Ken Sato (2017)

  5. No Access

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    Probing the Dynamics of Plasma Membrane Glutamate Transporters in Real Time by Total Internal Fluorescence Reflection Microscopy

    Plasma membrane solute carriers (SLCs) mediate the transport of a variety of substrates across the plasma membranes. There is a great interest in understanding how they work and how their density at the plasma...

    Eliana S. Di Cairano, Paola Marciani, Stefania Moretti, Carla Perego (2017)

  6. No Access

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    Induction of PINK1/Parkin-Mediated Mitophagy

    PINK1/Parkin mitophagy is a key mechanism to contribute mitochondrial quality control, and the defects are thought to be a cause of those Parkinson’s disease onsets. Upon loss of mitochondrial membrane potenti...

    Shigeto Sato, Norihiko Furuya (2017)

  7. No Access

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    A Simple Method of Generating 3D Brain Organoids Using Standard Laboratory Equipment

    3D brain organoids are a powerful tool with prospective application for the study of neural development and disease. Here we describe the growth factor-free method of generating cerebral organoids from feeder-...

    Magdalena Sutcliffe, Madeline A. Lancaster (2017)

  8. No Access

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    A Cleanup Method for Mass Spectrometric Analysis of Sphingosine- and Ceramide-1-Phosphate in Blood and Solid Tissue Using a Phosphate Capture Molecule

    Cleanup technology and mass spectrometric determination of sphingosine-1-phosphate (S1P) using a phosphate capture molecule are shown. The protocol is rapid, requires neither thin-layer chromatography nor liqu...

    Jun-ichi Morishige, Ryouhei Yamashita, Tamotsu Tanaka, Kiyoshi Satouchi (2017)

  9. No Access

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    The Use of Correlative Light-Electron Microscopy (CLEM) to Study PINK1/Parkin-Mediated Mitophagy

    In this chapter we describe the use of correlative light-electron microscopy (CLEM) to study, in cultured cells, the turnover of damaged mitochondria by PINK1/Parkin-dependent mitophagy. CLEM combines the adva...

    Chieko Kishi-Itakura, Folma Buss (2017)

  10. No Access

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    Human Intestinal Enteroids: New Models to Study Gastrointestinal Virus Infections

    Human rotavirus (HRV) and human norovirus (HuNoV) infections are recognized as the most common causes of epidemic and sporadic cases of gastroenteritis worldwide. The study of these two human gastrointestinal ...

    Winnie Y. Zou, Sarah E. Blutt, Sue E. Crawford, Khalil Ettayebi, Xi-Lei Zeng (2017)

  11. No Access

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    Immunocytochemical Monitoring of PINK1/Parkin-Mediated Mitophagy in Cultured Cells

    Both PINK1 and parkin are the responsible genes (PARK6 and PARK2, respectively) for familial early-onset Parkinson’s disease (PD). Several lines of evidences have suggested that mitochondrial dysfunction would...

    Motoki Fujimaki, Shinji Saiki, Yukiko Sasazawa, Kei-Ichi Ishikawa, Yoko Imamichi (2017)

  12. No Access

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    Sphingosine-1-Phosphate (S1P) Signaling in Neural Progenitors

    Sphingosine-1-phosphate (S1P) and its receptors are important in nervous system development. Reliable in vitro human model systems are needed to further define specific roles for S1P signaling in neural develo...

    Phillip Callihan, Mohammed Alqinyah, Shelley B. Hooks (2017)

  13. No Access

    Protocol

    Measuring Sphingosine-1-Phosphate: Protein Interactions with the Kinetic Exclusion Assay

    By directly detecting the ligand-free binding sites in a sample, the kinetic exclusion assay (KinExA®) provides a compelling alternative to SPR-based techniques for determining equilibrium dissociation constants ...

    Jonathan K. Fleming, Jonathan M. Wojciak (2017)

  14. No Access

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    Investigation of Yeast Mitophagy with Fluorescence Microscopy and Western Blotting

    Selective clearance of superfluous or dysfunctional mitochondria is a fundamental process that depends on the autophagic membrane trafficking pathways found in many cell types. This catabolic event, called mit...

    Sachiyo Nagumo, Koji Okamoto (2017)

  15. No Access

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    Exploring MicroRNAs on NIX-Dependent Mitophagy

    The dysregulation of autophagy is implicated in many pathological disorders including infections, aging, neurodegenerative diseases, and cancer. Autophagy can be precisely controlled both transcriptionally and...

    Wen Li, Hao Chen, Shupeng Li, Guanghong Lin, Du Feng (2017)

  16. No Access

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    Intestinal Crypt Organoid: Isolation of Intestinal Stem Cells, In Vitro Culture, and Optical Observation

    The isolation and culture of intestinal stem cells (ISCs) was first demonstrated in the very recent decade with the identification of ISC marker Lgr5. The growth of ISCs into crypt organoids provides an in vit...

    Yun Chen, Chuan Li, Ya-Hui Tsai, Sheng-Hong Tseng (2017)

  17. No Access

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    Quantum Dot Toolbox in Membrane Neurotransmitter Transporter Research

    Quantum dot-based fluorescence techniques enable multi-scale molecular profiling ranging from real-time single molecule dynamics to expression trends in million-cell populations. In comparison to currently ava...

    Lucas B. Thal, Danielle M. Bailey, Oleg Kovtun, Sandra J. Rosenthal (2017)

  18. No Access

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    Monitoring Mitochondrial Changes by Alteration of the PINK1-Parkin Signaling in Drosophila

    Mitochondrial quality control is a key process in tissues with high energy demands, such as the brain and muscles. Recent studies using Drosophila have revealed that the genes responsible for familial forms of ju...

    Tsuyoshi Inoshita, Kahori Shiba-Fukushima, Hongrui Meng, Nobutaka Hattori, Yuzuru Imai (2017)

  19. No Access

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    Mitophagy in Yeast: A Screen of Mitophagy-Deficient Mutants

    Mitochondrial autophagy (mitophagy) is a process that selectively degrades mitochondria via autophagy. Recent studies have shown that mitophagy plays an important role in mitochondrial homeostasis by degrading...

    Kentaro Furukawa, Tomotake Kanki (2017)

  20. No Access

    Protocol

    Erratum to: Cultivation and Preservation of Hydrocarbonoclastic Microorganisms, Particularly Cycloclasticus Species

    Maria Genovese, Renata Denaro, Daniela Russo, Francesca Crisafi, Santina Santisi (2017)

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