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A Robust Mammary Organoid System to Model Lactation and Involution-like Processes

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AIMTOR, a BRET Biosensor for Live Recording of mTOR Activity in Cell Populations and Single Cells

Authors: Nathalie Bouquier, Enora Moutin, Julie Perroy and Vincent Ollendorff, date: 04/20/2021, view: 32, Q&A: 0

[Abstract]

Mammalian target of rapamycin (mTOR) controls many crucial cellular functions, including protein synthesis, cell size, energy metabolism, lysosome and mitochondria biogenesis, and autophagy. Consequently, deregulation of mTOR signaling plays a role in numerous pathological conditions such as cancer, metabolic disorders and neurological diseases.

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A Versatile Protocol to Quantify BCR-mediated Phosphorylation in Human and Murine B Cell Subpopulations

Authors: Jasper Rip, Rudi W. Hendriks and Odilia B. J. Corneth, date: 02/05/2021, view: 492, Q&A: 0

[Abstract]

Signal transduction is the process by which molecular signals are transmitted from the cell surface to its interior, resulting in functional changes inside the cell. B cell receptor (BCR) signaling is of crucial importance for B cells, as it regulates their differentiation, selection, survival, cellular activation and proliferation. Upon BCR

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In vitro STING Activation with the cGAMP-STINGΔTM Signaling Complex

Authors: Yanpu He, Celestine Hong, Darrell J. Irvine, Jiahe Li and Paula T. Hammond, date: 02/05/2021, view: 615, Q&A: 0

[Abstract] Activating the STING (stimulator of interferon genes) signaling pathway via administration of STING agonist cyclic GMP-AMP (cGAMP) has shown great promise in cancer immunotherapy. While state-of-the-art approaches have predominantly focused on the encapsulation of cGAMP into liposomes or polymersomes for cellular delivery, we discovered that the ...

Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva

Authors: Raffaella Klima, Giulia Romano, Monsurat Gbadamosi, Aram Megighian and Fabian Feiguin, date: 02/05/2021, view: 382, Q&A: 0

[Abstract]

Alterations in synaptic transmission are critical early events in neuromuscular disorders. However, reliable methodologies to analyze the functional organization of the neuromuscular synapses are still needed. This manuscript provides a detailed protocol to analyze the molecular assembly of the neuromuscular synapses through

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Quantification of the Surface Expression of G Protein-coupled Receptors Using Intact Live-cell Radioligand Binding Assays

Authors: Xin Xu and Guangyu Wu, date: 09/20/2020, view: 1746, Q&A: 0

[Abstract] G protein-coupled receptors (GPCRs) are the most structurally diverse family of signaling proteins and regulate a variety of cell function. For most GPCRs, the cell surface is their functional destination where they are able to respond a wide range of extracellular stimuli, leading to the activation of intracellular signal transduction cascades. ...

Production and Bioassay of a Diffusible Factor that Induces Gametophyte-to-Sporophyte Developmental Reprogramming in the Brown Alga Ectocarpus

Authors: Haiqin Yao, Delphine Scornet, Yacine Badis, Akira F. Peters, Murielle Jam, Cecile Hervé, Philippe Potin, Susana M. Coelho and J. Mark Cock, date: 09/20/2020, view: 1631, Q&A: 0

[Abstract] The brown alga Ectocarpus has a haploid-diploid life cycle that involves alternation between two multicellular generations, the sporophyte and the gametophyte. Life cycle generation is not determined by ploidy but by a genetic system that includes two different three amino acid loop extension homeodomain transcription factors called ...

Flow-cytometric Detection of Low-level Reactive Oxygen Species in Cell Lines and Primary Immune Cells

Authors: Kevin Bode, Corinna Link, Peter H. Krammer and Heiko Weyd, date: 09/05/2020, view: 2455, Q&A: 0

[Abstract] Depending on its concentration and cellular origin the production of reactive oxygen species (ROS) in the organism serves a variety of functions. While high concentrations during an oxidative burst are used to fight pathogens, low to moderate amounts of ROS act as signaling molecules important for several physiological processes such as regulation ...

Assessing Gα_q/15-signaling with IP-One: Single Plate Transfection and Assay Protocol for Cell-Based High-Throughput Assay

Authors: Élie Besserer-Offroy, Rebecca L Brouillette, Jean-Michel Longpré and Philippe Sarret, date: 08/20/2020, view: 1342, Q&A: 0

[Abstract] Cell-based functional assays are an important part of compound screening and drug lead optimization, and they can also play a crucial role in the determination of the residues involved in ligand binding and signaling for a particular G-protein-coupled receptor. Conventional methods used for Gα_q/15-coupled receptors rely on the use of ...

Organotypic Slice Culture of the Embryonic Mouse Brain

Authors: James M. Clegg and Thomas Pratt, date: 07/05/2020, view: 1204, Q&A: 0

[Abstract] Organotypic slice culture is a powerful technique for exploring the embryonic development of the mammalian brain. In this protocol we describe a basic slice culture technique we have used for two sets of experiments: axon guidance transplant assays and bead culture assays.

Quantification of Protein Kinase A (PKA) Activity by An in vitro Radioactive Assay Using the Mouse Sperm Derived Enzyme

Authors: Cintia Stival, Carolina Baro Graf, Pablo E. Visconti and Dario Krapf, date: 06/20/2020, view: 1761, Q&A: 0

[Abstract] In order to acquire fertilizing potential, mammalian sperm must undergo a process known as capacitation, which relies on the early activation of Protein Kinase A (PKA). Frequently, PKA activity is assessed in whole-cell experiments by analyzing the phosphorylation status of its substrates in a western-blot. This technique ...

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