1,630 results match your criteria correlative light

Structural basis for VIPP1 oligomerization and maintenance of thylakoid membrane integrity.

Cell 2021 Jul 23;184(14):3643-3659.e23. Epub 2021 Jun 23.

Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany; Helmholtz Pioneer Campus, Helmholtz Zentrum München, 85764 Neuherberg, Germany; Department of Chemistry, Technical University of Munich, 85748 Garching, Germany. Electronic address:

Vesicle-inducing protein in plastids 1 (VIPP1) is essential for the biogenesis and maintenance of thylakoid membranes, which transform light into life. However, it is unknown how VIPP1 performs its vital membrane-remodeling functions. Here, we use cryo-electron microscopy to determine structures of cyanobacterial VIPP1 rings, revealing how VIPP1 monomers flex and interweave to form basket-like assemblies of different symmetries. Read More

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Where in the cell is my protein?

David J DeRosier

Q Rev Biophys 2021 06 21;54:e9. Epub 2021 Jun 21.

Brandeis University, Waltham, MA, USA.

The application of cryo-correlative light and cryo-electron microscopy (cryo-CLEM) gives us a way to locate structures of interest in the electron microscope. In brief, the structures of interest are fluorescently tagged, and images from the cryo-fluorescent microscope (cryo-FM) maps are superimposed on those from the cryo-electron microscope (cryo-EM). By enhancing cryo-FM to include single-molecule localization microscopy (SMLM), we can achieve much better localization. Read More

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Reactive oxygen FIB spin milling enables correlative workflow for 3D super-resolution light microscopy and serial FIB/SEM of cultured cells.

Sci Rep 2021 Jun 23;11(1):13162. Epub 2021 Jun 23.

Thermo Fisher Scientific, 5350 NE Dawson Creek Drive, Hillsboro, OR, 97124, USA.

Correlative light and electron microscopy (CLEM) is a powerful tool for defining the ultrastructural context of molecularly-labeled biological specimens, particularly when superresolution fluorescence microscopy (SRM) is used for CLEM. Current CLEM, however, is limited by the stark differences in sample preparation requirements between the two modalities. For CLEM using SRM, the small region of interest (ROI) of either or both modalities also leads to low success rate and imaging throughput. Read More

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Intracellular localisation of Mycobacterium tuberculosis affects efficacy of the antibiotic pyrazinamide.

Nat Commun 2021 06 21;12(1):3816. Epub 2021 Jun 21.

Host-Pathogen Interactions in Tuberculosis Laboratory, The Francis Crick Institute, London, UK.

To be effective, chemotherapy against tuberculosis (TB) must kill the intracellular population of the pathogen, Mycobacterium tuberculosis. However, how host cell microenvironments affect antibiotic accumulation and efficacy remains unclear. Here, we use correlative light, electron, and ion microscopy to investigate how various microenvironments within human macrophages affect the activity of pyrazinamide (PZA), a key antibiotic against TB. Read More

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Single cell analyses reveal distinct adaptation of typhoidal and non-typhoidal Salmonella enterica serovars to intracellular lifestyle.

PLoS Pathog 2021 Jun 18;17(6):e1009319. Epub 2021 Jun 18.

Abt. Mikrobiologie, Universität Osnabrück, Osnabrück, Germany.

Salmonella enterica is a common foodborne, facultative intracellular enteropathogen. Human-restricted typhoidal S. enterica serovars Typhi (STY) or Paratyphi A (SPA) cause severe typhoid or paratyphoid fever, while many S. Read More

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Probing morphology and chemistry in complex soft materials withresonant soft x-ray scattering.

J Phys Condens Matter 2021 Jun 17;33(31). Epub 2021 Jun 17.

Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States of America.

Small angle scattering methodologies have been evolving at fast pace over the past few decades due to the ever-increasing demands for more details on the complex nanostructures of multiphase and multicomponent soft materials like polymer assemblies and biomaterials. Currently, element-specific and contrast variation techniques such as resonant (elastic) soft/tender x-ray scattering, anomalous small angle x-ray scattering, and contrast-matching small angle neutron scattering, or combinations of above are routinely used to extract the chemical composition and spatial arrangement of constituent elements at multiple length scales and examine electronic ordering phenomena. Here we present some recent advances in selectively characterizing structural architectures of complex soft materials, which often contain multi-components with a wide range of length scales and multiple functionalities, where novel resonant scattering approaches have been demonstrated to decipher a higher level of structural complexity that correlates to functionality. Read More

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Correlative Super-Resolution and Atomic Force Microscopy of DNA Nanostructures and Characterization of Addressable Site Defects.

ACS Nano 2021 Jun 17. Epub 2021 Jun 17.

Center for Bio/Molecular Science and Engineering, U.S. Naval Research Laboratory Code 6900, Washington, D.C. 20375, United States.

To bring real-world applications of DNA nanostructures to fruition, advanced microscopy techniques are needed to shed light on factors limiting the availability of addressable sites. Correlative microscopy, where two or more microscopies are combined to characterize the same sample, is an approach to overcome the limitations of individual techniques, yet it has seen limited use for DNA nanotechnology. We have developed an accessible strategy for high resolution, correlative DNA-based points accumulation for imaging in nanoscale topography (DNA-PAINT) super-resolution and atomic force microscopy (AFM) of DNA nanostructures, enabled by a simple and robust method to selectively bind DNA origami to cover glass. Read More

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Super-resolution label-free volumetric vibrational imaging.

Nat Commun 2021 06 15;12(1):3648. Epub 2021 Jun 15.

Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, USA.

Innovations in high-resolution optical imaging have allowed visualization of nanoscale biological structures and connections. However, super-resolution fluorescence techniques, including both optics-oriented and sample-expansion based, are limited in quantification and throughput especially in tissues from photobleaching or quenching of the fluorophores, and low-efficiency or non-uniform delivery of the probes. Here, we report a general sample-expansion vibrational imaging strategy, termed VISTA, for scalable label-free high-resolution interrogations of protein-rich biological structures with resolution down to 78 nm. Read More

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Correlating Super-Resolution Microscopy and Transmission Electron Microscopy Reveals Multiparametric Heterogeneity in Nanoparticles.

Nano Lett 2021 06 14;21(12):5360-5368. Epub 2021 Jun 14.

Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Baldiri Reixac 15-21, 08028 Barcelona, Spain.

The functionalization of nanoparticles with functional moieties is a key strategy to achieve cell targeting in nanomedicine. The interplay between size and ligand number is crucial for the formulation performance and needs to be properly characterized to understand nanoparticle structure-activity relations. However, there is a lack of methods able to measure both size and ligand number at the same time and at the single particle level. Read More

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Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells.

J Vis Exp 2021 05 28(171). Epub 2021 May 28.

Harwell Science and Innovation Campus, Beamline B24, Diamond Light Source;

Three-dimensional (3D) structured illumination microscopy (SIM) allows imaging of fluorescently labelled cellular structures at higher resolution than conventional fluorescence microscopy. This super-resolution (SR) technique enables visualization of molecular processes in whole cells and has the potential to be used in conjunction with electron microscopy and X-ray tomography to correlate structural and functional information. A SIM microscope for cryogenically preserved samples (cryoSIM) has recently been commissioned at the correlative cryo-imaging beamline B24 at the UK synchrotron. Read More

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Thioesterase superfamily member 1 undergoes stimulus-coupled conformational reorganization to regulate metabolism in mice.

Nat Commun 2021 06 9;12(1):3493. Epub 2021 Jun 9.

Division of General Surgery, Department of Surgery, Beth Israel Deaconess Medical Center, Boston, MA, USA.

In brown adipose tissue, thermogenesis is suppressed by thioesterase superfamily member 1 (Them1), a long chain fatty acyl-CoA thioesterase. Them1 is highly upregulated by cold ambient temperature, where it reduces fatty acid availability and limits thermogenesis. Here, we show that Them1 regulates metabolism by undergoing conformational changes in response to β-adrenergic stimulation that alter Them1 intracellular distribution. Read More

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Cellular uptake and targeting of low dispersity, dual emissive, segmented block copolymer nanofibers.

Chem Sci 2020 Jul 8;11(32):8394-8408. Epub 2020 Jul 8.

School of Chemistry, University of Bristol Bristol BS8 1TS UK.

Polymer-based nanoparticles show substantial promise in the treatment and diagnosis of cancer and other diseases. Herein we report an exploration of the cellular uptake of tailored, low dispersity segmented 1D nanoparticles which were prepared from an amphiphilic block copolymer, poly(dihexylfluorene)--poly(ethyleneglycol) (PDHF--PEG), with a crystallizable PDHF core-forming block and a 'stealth' PEG corona-forming block with different end-group functionalities. Segmented C-B-A-B-C pentablock 1D nanofibers with varied spatially-defined coronal chemistries and a selected length (95 nm) were prepared using the living crystallization-driven self-assembly (CDSA) seeded-growth method. Read More

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Age influences susceptibility of brain capillary endothelial cells to La Crosse virus infection and cell death.

J Neuroinflammation 2021 Jun 3;18(1):125. Epub 2021 Jun 3.

Research and Technologies Branch, Rocky Mountain Laboratories, NIAID, NIH, 903 S. 4th Street, MT, 59840, Hamilton, USA.

Background: A key factor in the development of viral encephalitis is a virus crossing the blood-brain barrier (BBB). We have previously shown that age-related susceptibility of mice to the La Crosse virus (LACV), the leading cause of pediatric arbovirus encephalitis in the USA, was associated with the ability of the virus to cross the BBB. LACV infection in weanling mice (aged around 3 weeks) results in vascular leakage in the olfactory bulb/tract (OB/OT) region of the brain, which is not observed in adult mice aged > 6-8 weeks. Read More

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Malar stripe size and prominence in peregrine falcons vary positively with solar radiation: support for the solar glare hypothesis.

Biol Lett 2021 06 2;17(6):20210116. Epub 2021 Jun 2.

FitzPatrick Institute of African Ornithology, DSI-NRF Centre of Excellence, Department of Biological Sciences, University of Cape Town, South Africa.

Many falcons ( spp.) exhibit a distinct dark plumage patch below the eye, termed the malar stripe. This stripe is hypothesized to reduce the amount of solar glare reflected into the eyes while foraging, thereby increasing hunting efficiency in bright conditions. Read More

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Protocol for image registration of correlative soft X-ray tomography and super-resolution structured illumination microscopy images.

STAR Protoc 2021 Jun 6;2(2):100529. Epub 2021 May 6.

Université de Nantes, CHU Nantes, CNRS UMR 6291, INSERM UMR 1087, L'institut du thorax, 44000 Nantes, France.

Correlation of 3D images acquired on different microscopes can be a daunting prospect even for experienced users. This protocol describes steps for registration of images from soft X-ray absorption contrast imaging and super-resolution fluorescence imaging of hydrated biological materials at cryogenic temperatures. Although it is developed for data generated at synchrotron beamlines that offer the above combination of microscopies, it is applicable to all analogous imaging systems where the same area of a sample is examined using successive non-destructive imaging techniques. Read More

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Unveiling a Hidden Event in Fluorescence Correlative Microscopy by AFM Nanomechanical Analysis.

Front Mol Biosci 2021 6;8:669361. Epub 2021 May 6.

Shenzhen Key Laboratory of Nanobiomechanics, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

Fluorescent imaging combined with atomic force microscopy (AFM), namely AFM-fluorescence correlative microscopy, is a popular technology in life science. However, the influence of involved fluorophores on obtained mechanical information is normally underestimated, and such subtle changes are still challenging to detect. Herein, we combined AFM with laser light excitation to perform a mechanical quantitative analysis of a model membrane system labeled with a commonly used fluorophore. Read More

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Increased teat wall thickness in response to machine milking.

J Dairy Sci 2021 Aug 21;104(8):9082-9092. Epub 2021 May 21.

Veterinary Physiology, Vetsuisse Faculty, University of Bern, 3012 Bern, Switzerland. Electronic address:

Transiently increased teat wall thickness in response to machine milking has been documented by various methods, including ultrasound. However, correlative ultrasonography and histology to detect the origin of this phenomenon is lacking. The first goal of the present study was to evaluate and compare milking-related changes of the teat tissue in 2 breeds of dairy cows (11 Simmental and 3 Holstein) using B-mode ultrasonography. Read More

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Interrater Reliability of NI-RADS on Posttreatment PET/Contrast-enhanced CT Scans in Head and Neck Squamous Cell Carcinoma.

Radiol Imaging Cancer 2021 05;3(3):e200131

From the Department of Radiology, Emory University School of Medicine, 1364 Clifton Rd NE, Room BG03, Atlanta, GA 30322 (D.H., A.H.A.); Department of Neuroradiology, Mayo Clinic, Phoenix, Ariz (T.J.R.); Departments of Radiology and Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, Pa (B.F.B.); Department of Radiology and Imaging Sciences, University of Utah Health Sciences Center, Salt Lake City, Utah (Y.A., R.H.W.); Department of Neuroradiology, Weill Cornell Imaging at New York-Presbyterian, New York, NY (C.D.P.); Department of Radiology, Massachusetts Eye and Ear, Harvard Medical School, Boston, Mass (A.F.J.); Department of Radiology and Imaging Sciences, Indiana University School of Medicine, Indianapolis, Ind (K.M.K.); Department of Radiology, University of Vermont Medical Center, Burlington, Vt (M.P.B.); Radiology Imaging Associates, Englewood, Colo (S.M.P.); Department of Radiology, Mayo Clinic, Jacksonville, Fla (M.H.K.); Department of Biostatistics and Bioinformatics, Emory University Rollins School of Public Health, Atlanta, Ga (P.A.R.); and Department of Radiology and Imaging Sciences, Emory University Hospital, Atlanta, Ga (B.R.).

Purpose To evaluate the interrater reliability among radiologists examining posttreatment head and neck squamous cell carcinoma (HNSCC) fluorodeoxyglucose PET/contrast-enhanced CT (CECT) scans using Neck Imaging Reporting and Data System (NI-RADS). Materials and Methods In this retrospective study, images in 80 patients with HNSCC who underwent posttreatment surveillance PET/CECT and immediate prior comparison CECT or PET/CECT (from June 2014 to July 2016) were uploaded to the American College of Radiology's cloud-based website, Cortex. Eight radiologists from seven institutions with variable NI-RADS experience independently evaluated each case and assigned an appropriate prose description and NI-RADS category for the primary site and the neck site. Read More

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Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures.

Nat Protoc 2021 06 14;16(6):2851-2885. Epub 2021 May 14.

Beamline B24, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, UK.

3D correlative microscopy methods have revolutionized biomedical research, allowing the acquisition of multidimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometer resolution imaging (2-50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super-resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample, resulting in the immediate requirement for protocols that facilitate hassle-free, unambiguous cross-correlation between microscopes. Read More

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Structural insight into transmissive mutant huntingtin species by correlative light and electron microscopy and cryo-electron tomography.

Biochem Biophys Res Commun 2021 06 10;560:99-104. Epub 2021 May 10.

Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA. Electronic address:

Aggregates of mutant huntingtin (mHTT) containing an expanded polyglutamine (polyQ) tract are hallmarks of Huntington's Disease (HD). Studies have shown that mHTT can spread between cells, leading to the propagation of misfolded protein pathology. However, the structure of transmissive mHTT species, and the molecular mechanisms underlying their transmission remain unknown. Read More

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Correlative fluorescence microscopy, transmission electron microscopy and secondary ion mass spectrometry (CLEM-SIMS) for cellular imaging.

PLoS One 2021 10;16(5):e0240768. Epub 2021 May 10.

Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.

Electron microscopy (EM) has been employed for decades to analyze cell structure. To also analyze the positions and functions of specific proteins, one typically relies on immuno-EM or on a correlation with fluorescence microscopy, in the form of correlated light and electron microscopy (CLEM). Nevertheless, neither of these procedures is able to also address the isotopic composition of cells. Read More

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Light and electron microscopy imaging unveils new aspects of the antiviral capacity of silver nanoparticles in bunyavirus-infected cells.

Virus Res 2021 Sep 4;302:198444. Epub 2021 May 4.

Cell Structure Laboratory, National Center for Biotechnology, CNB-CSIC, Campus UAM, Cantoblanco, 28049, Madrid, Spain. Electronic address:

Drug repurposing is an important source of new antivirals because many compounds used to treat a variety of pathologies also hamper viral infections. Habitually, silver nanoparticles (AgNPs) have been used to treat bacterial and fungal infections and their antiviral properties have been also reported. In this work, we have studied the antiviral capacity of AgNPs in cells infected with Bunyamwera virus (BUNV), the prototype of the Bunyavirales order. Read More

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September 2021

Correlative operando microscopy of oxygen evolution electrocatalysts.

Nature 2021 05 5;593(7857):67-73. Epub 2021 May 5.

Department of Materials Science and Engineering, Stanford University, Stanford, CA, USA.

Transition metal (oxy)hydroxides are promising electrocatalysts for the oxygen evolution reaction. The properties of these materials evolve dynamically and heterogeneously with applied voltage through ion insertion redox reactions, converting materials that are inactive under open circuit conditions into active electrocatalysts during operation. The catalytic state is thus inherently far from equilibrium, which complicates its direct observation. Read More

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Recapitulating Actin Module Organization in the Oocyte Reveals New Roles for Bristle-Actin-Modulating Proteins.

Int J Mol Sci 2021 Apr 13;22(8). Epub 2021 Apr 13.

Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel.

The generation of F-actin bundles is controlled by the action of actin-binding proteins. In bristle development, two major actin-bundling proteins-Forked and Fascin-were identified, but still the molecular mechanism by which these actin-bundling proteins and other proteins generate bristle actin bundles is unknown. In this study, we developed a technique that allows recapitulation of bristle actin module organization using the ovary by a combination of confocal microscopy, super-resolution structured illumination microscopy, and correlative light and electron microscope analysis. Read More

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Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy.

Int J Mol Sci 2021 Apr 20;22(8). Epub 2021 Apr 20.

Integrative Research Support Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.

Immuno-electron microscopy (Immuno-EM) is a powerful tool for identifying molecular targets with ultrastructural details in biological specimens. However, technical barriers, such as the loss of ultrastructural integrity, the decrease in antigenicity, or artifacts in the handling process, hinder the widespread use of the technique by biomedical researchers. We developed a method to overcome such challenges by combining light and electron microscopy with immunolabeling based on Tokuyasu's method. Read More

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Microscopic Observation of SARS-Like Particles in RT-qPCR SARS-CoV-2 Positive Sewage Samples.

Pathogens 2021 Apr 24;10(5). Epub 2021 Apr 24.

Microbes, Evolution, Phylogeny and Infection (MEPHI), UM63, Institut de Recherche pour le Développement (IRD), Aix- Marseille University, 13005 Marseille, France.

The ongoing outbreak of novel coronavirus pneumonia (COVID-19) caused by SARS-CoV-2 infection has spread rapidly worldwide. The major transmission routes of SARS-CoV-2 are recognised as inhalation of aerosol/droplets and person-to-person contact. However, some studies have demonstrated that live SARS-CoV-2 can be isolated from the faeces and urine of infected patients, which can then enter the wastewater system. Read More

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HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells.

Elife 2021 Apr 27;10. Epub 2021 Apr 27.

Department of Infectious Diseases Virology, University Hospital Heidelberg, Heidelberg, Germany.

HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization, and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed HIV-1 cDNA molecules and their association with viral and cellular proteins using fluorescence and correlative-light-and-electron-microscopy (CLEM). We specifically detected HIV-1 cDNA inside nuclei, but not in the cytoplasm. Read More

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Antigen-independent activation enhances the efficacy of 4-1BB-costimulated CD22 CAR T cells.

Nat Med 2021 05 22;27(5):842-850. Epub 2021 Apr 22.

Center for Cellular Immunotherapies, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.

While CD19-directed chimeric antigen receptor (CAR) T cells can induce remission in patients with B cell acute lymphoblastic leukemia (ALL), a large subset relapse with CD19 disease. Like CD19, CD22 is broadly expressed by B-lineage cells and thus serves as an alternative immunotherapy target in ALL. Here we present the composite outcomes of two pilot clinical trials ( NCT02588456 and NCT02650414 ) of T cells bearing a 4-1BB-based, CD22-targeting CAR in patients with relapsed or refractory ALL. Read More

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Membranous Structures Directly Come in Contact With p62/SQSTM1 Bodies.

J Histochem Cytochem 2021 Jun 22;69(6):407-414. Epub 2021 Apr 22.

Department of Cellular and Molecular Neuropathology, Graduate School of Medicine, Juntendo University, Tokyo, Japan.

During autophagy, autophagosomes are formed to engulf cytoplasmic contents. p62/SQSTM-1 is an autophagic adaptor protein that forms p62 bodies. A unique feature of p62 bodies is that they seem to directly associate with membranous structures. Read More

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