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Protein-based, fluorescent biosensors power basic research on cell signaling in health and disease, but their use in automated laboratories is limited. We have now created two live-cell assays, one for diacyl glycerol and another for cAMP, that are robust (Z' > 0.7) and easily deployed on standard fluorescence plate readers. We describe the development of these assays, focusing on the parameters that were critical for optimization, in the hopes that the lessons learned can be generalized to the development of new biosensor-based assays.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4766961 | PMC |
http://dx.doi.org/10.1177/1087057115618608 | DOI Listing |
J Virol
September 2025
Genome Regulation and Cell Signaling, Ellen and Ronald Caplan Cancer Center, The Wistar Institute, Philadelphia, Pennsylvania, USA.
Unlabelled: Adenoviruses are double-stranded DNA viruses widely used as platforms for vaccines, oncolytics, and gene delivery. However, tools for studying adenoviral gene expression in real time during infection remain limited. Here, we describe a set of fluorescent and bioluminescent reporter viruses built using the modular AdenoBuilder reverse genetics system and informed by high-resolution maps of Ad5 transcription.
View Article and Find Full Text PDFFront Immunol
September 2025
Department of Psychiatry and Psychotherapy, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Introduction: Anti-N-methyl-D-aspartate receptor (NMDA-R) encephalitis is a neuropsychiatric disorder with additional psychiatric features caused by NMDA-R immunoglobulin G (IgG) antibodies in cerebrospinal fluid (CSF). This report presents the follow-up of a patient in whom we assumed mild NMDA-R encephalitis in the first psychotic episode.
Case Study: A patient with a prior episode of an acute polymorphic psychotic syndrome relapsed five and a half years later following a severe COVID-19 infection.
Redox Biol
September 2025
Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong First People's Hospital, Medical School of Nantong University, Nantong, Jiangsu, 226000, China; Research Institute for Spine and Spinal Cord Disease of Nantong University, Nantong, Jiangsu, 226000, China. Elec
Spinal cord injury (SCI) is a devastating condition characterized by the accumulation of myelin debris (MD), persistent neuroinflammation, and impaired neural regeneration. Although macrophages are pivotal for MD clearance, the impact of excessive MD phagocytosis on macrophage phenotype and function remains poorly understood. Building upon our prior evidence that exendin-4 (Ex-4), a glucagon-like peptide-1 receptor (GLP-1R) agonist, mitigates microglia-driven neuroinflammation post-SCI, this study elucidates the therapeutic efficacy and underlying mechanisms of Ex-4 in alleviating macrophage senescence, restoring efferocytotic capacity, and facilitating neural repair.
View Article and Find Full Text PDFCell Rep
September 2025
Department of Biochemistry, University of Colorado, Boulder, CO 80303, USA. Electronic address:
RNA polymerase II (RNAPII) is regulated by sequence-specific transcription factors (TFs) and the pre-initiation complex (PIC): TFIIA, TFIIB, TFIID, TFIIE, TFIIF, TFIIH, and Mediator. TFs, Mediator, and RNAPII contain intrinsically disordered regions (IDRs) and form phase-separated condensates, but how IDRs control RNAPII function remains poorly understood. Using purified PIC factors, we developed a real-time in vitro fluorescence transcription (RIFT) assay for second-by-second visualization of transcription at hundreds of promoters simultaneously.
View Article and Find Full Text PDFSTAR Protoc
September 2025
Division of Pulmonary Medicine and Critical Care, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA; Department of Systems Biology and Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA; Department of Pulmonary Medicine, Cincinnati Children's Hospital Medical C
Calcium signaling is crucial for endothelial cell homeostasis. Alterations in intracellular calcium levels due to shear stress are linked to vascular dysfunction and diseases. Here, we present a protocol to perform live calcium imaging by using a live calcium indicator on human lung endothelial cells subjected to shear stress in a commercially available microfluidic device (Ibidi Luer VI).
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