Cell Biology

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    Protocols in Current Issue
    14C-paraquat Efflux Assay in Arabidopsis Mesophyll Protoplasts
    Authors:  Jin-Qiu Xia, Qian-Qian Liu and Cheng-Bin Xiang, date: 09/20/2022, view: 78, Q&A: 0
    [Abstract]

    Weeds compete with crops for growth resources, causing tremendous yield losses. Paraquat is one of the three most common non-selective herbicides. To study the mechanisms of paraquat resistance, we need to trace the movement of paraquat in plants and within the cell. 14C is a radioactive carbon isotope widely used to trace substances of

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    In vivo Characterization of Endogenous Protein Interactomes in Drosophila Larval Brain, Using a CRISPR/Cas9-based Strategy and BioID-based Proximity Labeling
    Authors:  Ezgi Uçkun, Georg Wolfstetter, Johannes Fuchs and Ruth H. Palmer, date: 07/05/2022, view: 1371, Q&A: 0
    [Abstract]

    Understanding protein-protein interactions (PPIs) and interactome networks is essential to reveal molecular mechanisms mediating various cellular processes. The most common method to study PPIs in vivo is affinity purification combined with mass spectrometry (AP–MS). Although AP–MS is a powerful method, loss of weak and transient

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    CRISPR/Cas9 Gene Editing of HeLa Cells to Tag Proteins with mNeonGreen
    Authors:  Sachin Surve and Alexander Sorkin, date: 05/20/2022, view: 1452, Q&A: 0
    [Abstract]

    Subcellular localization dynamics of proteins involved in signal transduction processes is crucial in determining the signaling outcome. However, there is very limited information about the localization of endogenous signaling proteins in living cells. For example, biochemical mechanisms underlying the signaling pathway from epidermal growth

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    Measurement of Cell Intrinsic TGF-β Activation Mediated by the Integrin αvβ8
    Authors:  Robert Ian Seed and Stephen Lloyd Nishimura, date: 04/20/2022, view: 1571, Q&A: 0
    [Abstract]

    Transforming growth factor beta (TGF-β) is a multi-functional cytokine that plays a significant role in multiple diseases, including fibrosis and tumor progression. Whilst the biologic effects of TGF-β are well characterized, it is unclear how TGF-β signaling is regulated to impart specific responses within certain cell types. One

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    Activation of Mitochondrial Ca2+ Oscillation and Mitophagy Induction by Femtosecond Laser Photostimulation
    Authors:  Xiaoying Tian and Hao He, date: 04/05/2022, view: 735, Q&A: 0
    [Abstract]

    Ultra-precise stimulation solely to individual mitochondria, without any influence to the whole cell, is quite difficult by traditional biochemical reagents. In mitophagy research, the mitochondria and even the whole cell usually suffer irreversible and great damage caused by treatment with potent chemicals. In this protocol, we present the

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    A Hypersensitive Genetically Encoded Fluorescent Indicator (roGFP2-Prx1) Enables Continuous Measurement of Intracellular H2O2 during Cell Micro-cultivation
    Authors:  Veronica Gast, Verena Siewers and Mikael Molin, date: 02/05/2022, view: 1485, Q&A: 0
    [Abstract]

    Hydrogen peroxide (H2O2) is a toxic oxidant produced as a byproduct of several biological processes. At too high levels of hydrogen peroxide cells will experience oxidative stress, leading to a cellular response to decrease its levels and to protect the cells. Previously, methods used to study and quantify intracellular H2O2 have been limited by

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    Chronic Daily House Dust Mite Exposure in Mice is an Effective Model to Quantify the Effect of Pharmacologic Agents on Discrete Stages of Artery Remodeling in Pulmonary Hypertension
    Authors:  Lea C. Steffes and Maya E. Kumar, date: 01/05/2022, view: 1788, Q&A: 0
    [Abstract]

    Pulmonary hypertension (PH) is a heterogenous and incurable disease marked by varying degrees of pulmonary vascular remodeling. This vascular remodeling, which includes thickening of the smooth muscle layer (an early finding) and formation of occlusive neointimal lesions (a late finding) in the pulmonary arteries, is a major driver of morbidity

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    In vitro Fluid Shear Stress Induced Sclerostin Degradation and CaMKII Activation in Osteocytes
    [Abstract]

    Bone is a dynamic tissue that adapts to changes in its mechanical environment. Mechanical stimuli pressurize interstitial fluid in the lacunar-canalicular system within the bone matrix, causing fluid shear stress (FSS) across bone embedded, mechano-sensitive osteocytes. Therefore, modeling this mechanical stimulus in vitro is vital for identifying

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    Modeling the Nonlinear Dynamics of Intracellular Signaling Networks
    Authors:  Oleksii S. Rukhlenko and Boris N. Kholodenko, date: 07/20/2021, view: 2381, Q&A: 0
    [Abstract]

    This protocol illustrates a pipeline for modeling the nonlinear behavior of intracellular signaling pathways. At fixed spatial points, nonlinear signaling dynamics are described by ordinary differential equations (ODEs). At constant parameters, these ODEs may have multiple attractors, such as multiple steady states or limit cycles. Standard

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    Ethanol-induced Sedative Behavior: An Assay to Investigate Increased Dopamine Signaling in Caenorhabditis elegans
    Authors:  Anuradha Singh, Kavita Babu and Pratima Pandey, date: 07/05/2021, view: 1970, Q&A: 0
    [Abstract]

    Dopamine (DA) signaling affects locomotion, feeding, learning, and memory in C. elegans. Various assays have been developed to study the proteins involved in these behaviors; however, these assays show behavioral output only when there is a drastic change in DA levels. We designed an assay capable of observing behavioral output even with only

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