Featured
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| Open AccessBirefringent Fourier filtering for single molecule coordinate and height super-resolution imaging with dithering and orientation
Determining the orientation of single molecules in super resolution imaging is challenging. Here, by adding polarization control to phase control in the Fourier plane of the imaging path, parameters such as 3D spatial position, 3D orientation and wobbling or dithering angle can be determined from single molecules.
- Valentina Curcio
- , Luis A. Alemán-Castañeda
- & Miguel A. Alonso
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Article
| Open AccessComplementary mesoscale dynamics of spectrin and acto-myosin shape membrane territories during mechanoresponse
The cell cortex that supports the plasma membrane contains spectrin, a protein that interacts with the acto-myosin cytoskeleton. Here, the authors analyze spectrin behavior during cellular mechanoresponse and membrane trafficking, and observe spectrin regulation by myosin-driven contractility.
- Andrea Ghisleni
- , Camilla Galli
- & Nils C. Gauthier
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Article
| Open AccessRaman-guided subcellular pharmaco-metabolomics for metastatic melanoma cells
Single-cell metabolomics can offer deep insights into the metabolic reprogramming that accompanies disease states. Here, the authors use Raman spectro-microscopy for non-invasive metabolite analysis and identification of druggable metabolic susceptibilities in single live melanoma cells.
- Jiajun Du
- , Yapeng Su
- & Lu Wei
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Article
| Open AccessT-Plastin reinforces membrane protrusions to bridge matrix gaps during cell migration
In vivo, cells migrate across a diverse landscape of extracellular matrix containing gaps which present a challenge for cells to protrude across. Here, the authors show that T-Plastin strengthens protrusive actin networks to promote protrusion, extracellular matrix gap-bridging, and cell migration.
- Damien Garbett
- , Anjali Bisaria
- & Tobias Meyer
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Article
| Open AccessUltralong purely organic aqueous phosphorescence supramolecular polymer for targeted tumor cell imaging
Room-temperature phosphorescence has attracted attention for bioimaging, but quenching in aqueous systems is an issue. Here, the authors report on the synthesis of a water soluble organic phosphorescent polymer and demonstrate application is cancer cell targeting and imaging.
- Wei-Lei Zhou
- , Yong Chen
- & Yu Liu
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Article
| Open AccessTwo-color nanoscopy of organelles for extended times with HIDE probes
High density environmentally sensitive (HIDE) probes allow for long time-lapse super-resolution imaging of live cells. Here the authors develop a second HIDE probe with a bio-orthogonal labelling strategy to enable two-color nanoscopy of two organelles over extended periods.
- Ling Chu
- , Jonathan Tyson
- & Derek K. Toomre
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Article
| Open AccessMutant p53 induces Golgi tubulo-vesiculation driving a prometastatic secretome
p53 mutants can promote tumorigenesis by affecting fundamental cellular pathways and functions. In this study, the authors demonstrate a novel mutant-p53/HIF1α/miR-30d axis that impacts Golgi structure, trafficking, and secretion of proteins essential for tumor growth and metastasis.
- Valeria Capaci
- , Lorenzo Bascetta
- & Giannino Del Sal
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Article
| Open AccessLight microscopy of proteins in their ultrastructural context
Imaging specific proteins in the ultrastructural context largely relies on correlative light/electron microscopy, but fluorophore incompatibility and registration issues limit its use. Here the authors develop an expansion microscopy method with pan-labeling of the proteome to obtain EM-equivalent light microscopy images.
- Ons M’Saad
- & Joerg Bewersdorf
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Article
| Open AccessIn vivo microscopy reveals macrophage polarization locally promotes coherent microtubule dynamics in migrating cancer cells
The regulation of microtubule (MT) dynamics in cancer cells within the tumor microenvironment is less understood. Here, the authors develop an imaging platform to examine MT dynamics in live xenograft models and show that pro-tumor macrophages modulate MT coherence and alignment to promote cancer cell migration.
- Gaurav Luthria
- , Ran Li
- & Miles A. Miller
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Article
| Open AccessMultiplexed imaging of nucleome architectures in single cells of mammalian tissue
The three-dimensional architecture of the genome affects genomic functions. Here, the authors developed Multiplexed Imaging of Nucleome Architectures to measure multiscale chromatin folding, RNA profiles, and associations of numerous genomic regions with nuclear lamina and nucleoli in the same, single cells in heterogeneous tissue.
- Miao Liu
- , Yanfang Lu
- & Siyuan Wang
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Article
| Open AccessSuper-resolution imaging reveals the evolution of higher-order chromatin folding in early carcinogenesis
Aberrant chromatin structure is often found in cancer. Here, the authors optimise super-resolution microscopy for pathological tissue and discovered a significant decompaction of chromatin folding in early carcinogenesis prior to tumour formation.
- Jianquan Xu
- , Hongqiang Ma
- & Yang Liu
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Article
| Open AccessImaging small molecule-induced endosomal escape of siRNA
Therapeutic siRNA becomes trapped in endosomes, limiting its efficacy. Here the authors use fluorescently-tagged galectin-9 as a biosensor for membrane damage to monitor endosomal escape of cholesterol-conjugated siRNA following treatment of small molecule membrane-destabilising drugs.
- Hampus Du Rietz
- , Hampus Hedlund
- & Anders Wittrup
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Article
| Open AccessAngiotensin II induces coordinated calcium bursts in aldosterone-producing adrenal rosettes
Aldosterone-producing zona glomerulosa cells in the adrenal gland arrange into rosette structures known to be important for morphogenesis. Here the authors show that the cells in the rosettes produce coordinated calcium activity bursts in response to angiotensin II that correlate with aldosterone production level.
- Nick A. Guagliardo
- , Peter M. Klein
- & Mark P. Beenhakker
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Article
| Open AccessParadoxical activation of the protein kinase-transcription factor ERK5 by ERK5 kinase inhibitors
Selective ERK5 inhibitors target ERK5 kinase activity, but they do not phenocopy the effects of ERK5 genetic depletion. Here, the authors demonstrate that the direct interaction of these inhibitors to ERK5 kinase domain induces conformational changes that promote ERK5 nuclear translocation and transcriptional activities.
- Pamela A. Lochhead
- , Julie A. Tucker
- & Simon J. Cook
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Article
| Open AccessPALB2 chromatin recruitment restores homologous recombination in BRCA1-deficient cells depleted of 53BP1
Although loss of BRCA1 leads to defects in DNA double-strand break repair by homologous recombination (HR) and renders cells hypersensitive to PARP inhibitors, resistance to the drugs can arise. Here the authors reveal that PALB2 chromatin recruitment restores HR in BRCA1-deficient cells depleted of 53BP1.
- Rimma Belotserkovskaya
- , Elisenda Raga Gil
- & Stephen P. Jackson
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Article
| Open AccessSpatiotemporal contact between peroxisomes and lipid droplets regulates fasting-induced lipolysis via PEX5
Lipid droplets are organelles that regulate lipid metabolism but if organellar contacts play a role during lipolysis is unclear. Here, the authors show that peroxisomes and peroxisomal protein PEX5 play pivotal roles in the spatial and temporal regulation of fasting-induced lipolysis by translocating ATGL onto lipid droplets
- Jinuk Kong
- , Yul Ji
- & Jae Bum Kim
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Article
| Open AccessSuper-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics
Glucagon-like peptide-1 receptor is an important regulator of appetite and glucose homeostasis. Here the authors describe super-resolution microscopy and in vivo imaging compatible fluorescent probes, which reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics in islets and brain.
- Julia Ast
- , Anastasia Arvaniti
- & David J. Hodson
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Article
| Open AccessA set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales
Monomeric near-infrared (NIR) fluorescent proteins (FPs) from bacterial phytochromes bring potential advantages, but their brightness in cells is lower than dimeric NIR FPs. Here the authors develop enhanced monomeric NIR FPs enabling imaging across different scales without the trade-off between brightness and monomeric state.
- Mikhail E. Matlashov
- , Daria M. Shcherbakova
- & Vladislav V. Verkhusha
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Article
| Open AccessMicroparticle traction force microscopy reveals subcellular force exertion patterns in immune cell–target interactions
Traction force microscopy is an effective method for measuring cellular forces but it is limited by planar geometry. Here the authors develop a facile method to produce deformable hydrogel particles and a reference-free computational method to resolve surface traction forces from particle shape deformation.
- Daan Vorselen
- , Yifan Wang
- & Julie A. Theriot
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Article
| Open AccessQuantitative live cell imaging reveals influenza virus manipulation of Rab11A transport through reduced dynein association
Here, using high spatiotemporal resolution light-sheet and fluorescence microscopy, the authors investigate the role of cytoskeletal components on the intracellular transport of Rab11A and influenza virus (IAV) vRNP), and show a preference for Rab11A movement along microtubules that is not essential for IAV vRNP transport.
- Amar R. Bhagwat
- , Valerie Le Sage
- & Seema S. Lakdawala
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Article
| Open AccessPhagocytosis is mediated by two-dimensional assemblies of the F-BAR protein GAS7
The Bin/Amphiphysin/Rvs167 (BAR) domain superfamily, which includes FCH-BAR (F-BAR) domain proteins are membrane-sculpting proteins. Here the authors combine a range of techniques and show that the F-BAR domain of growth-arrest specific protein 7 (GAS7) forms two-dimensional sheets on flat membranes and that these oligomeric assemblies of GAS7 are required for the formation of phagocytic cups in macrophages.
- Kyoko Hanawa-Suetsugu
- , Yuzuru Itoh
- & Shiro Suetsugu
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Article
| Open AccessSuperresolution architecture of cornerstone focal adhesions in human pluripotent stem cells
Pluripotent stem cell colonies are encircled by large cornerstone focal adhesions (FAs). Here, using super-resolution imaging, the authors describe features in the nanoscale makeup of these stable FAs such as inverted vinculin, lateral talin segregation and distinct kank protein distributions.
- Aki Stubb
- , Camilo Guzmán
- & Johanna Ivaska
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Article
| Open AccessThe ALFA-tag is a highly versatile tool for nanobody-based bioscience applications
Epitope tags are widely used in various applications, but often lack versatility. Here, the authors introduce a small, alpha helical tag, which is recognized by a high affinity nanobody and can be used in a range of different applications, from protein purification to super-resolution imaging and in vivo detection of proteins.
- Hansjörg Götzke
- , Markus Kilisch
- & Steffen Frey
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Article
| Open AccessMiro clusters regulate ER-mitochondria contact sites and link cristae organization to the mitochondrial transport machinery
Mitochondrial cristae organization and ER-mitochondria contact sites are critical structures for cellular function. Here, the authors use super-resolution microscopy to show that Miro GTPases form clusters required for normal ER-mitochondria contact sites formation and to link cristae organization to the mitochondrial transport machinery.
- Souvik Modi
- , Guillermo López-Doménech
- & Josef T. Kittler
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Article
| Open AccessFlotillins promote T cell receptor sorting through a fast Rab5–Rab11 endocytic recycling axis
Internalized receptors are recycled back to the cell surface, but their precise mechanisms are unclear. Here, the authors show that the flotillin membrane proteins may regulate the transfer of internalized T cell receptors into Rab5 and Rab11-positive endosomes to support its rapid recycling.
- Gregory M. I. Redpath
- , Manuela Ecker
- & Jérémie Rossy
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Article
| Open AccessA highly dynamic F-actin network regulates transport and recycling of micronemes in Toxoplasma gondii vacuoles
Replication of Toxoplasma gondii requires replication and distribution of essential organelles such as micronemes. Here, Periz et al. show that micronemes are recycled from the mother to the forming daughter cells using a highly dynamic F-actin network that supports multidirectional vesicle transport.
- Javier Periz
- , Mario Del Rosario
- & Markus Meissner
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Article
| Open AccessStructural basis for delta cell paracrine regulation in pancreatic islets
Pancreatic islets are composed of alpha-, beta-, as well as delta-cells and appropriate regulation of glucose homeostasis relies on auto- and paracrine cellular communication. Here, the authors study the role of delta-cell filopodia in this context by employing optogenetic and calcium imaging approaches.
- Rafael Arrojo e Drigo
- , Stefan Jacob
- & Per-Olof Berggren
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Article
| Open AccessStudying nucleic envelope and plasma membrane mechanics of eukaryotic cells using confocal reflectance interferometric microscopy
Biomechanical studies of eukaryotic cells have been limited due to low sensitivity and axial resolution in interferometric imaging. Here, the authors present depth-resolved confocal reflectance interferometric microscopy with high sensitivity and temporal resolution, which enables quantification of nucleic envelope and plasma membrane fluctuations.
- Vijay Raj Singh
- , Yi An Yang
- & Peter T. C. So
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Article
| Open AccessDegron-tagged reporters probe membrane topology and enable the specific labelling of membrane-wrapped structures
Visualising certain organelles and their dynamics is challenging in living cells. Here the authors co-opt selective degradation to label membrane-bound compartments in worm embryos and mammalian cells, revealing membrane topology during cell division.
- Katharina B. Beer
- , Gholamreza Fazeli
- & Ann M. Wehman
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Article
| Open AccessSingle-molecule localization microscopy and tracking with red-shifted states of conventional BODIPY conjugates in living cells
Single-molecule localization microscopy (SMLM) requires the use of fluorophores with specific sets of properties. Here the authors employ conventional BODIPY dyes as SMLM fluorophores by making use of rarely reported red-shifted ground state BODIPY dimers to image fatty acids, lipid droplets and lysosomes at single-molecule resolution.
- Santosh Adhikari
- , Joe Moscatelli
- & Elias M. Puchner
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Article
| Open AccessTwo-step chromosome segregation in the stalked budding bacterium Hyphomonas neptunium
In bacteria, DNA replication and segregation are commonly coupled. Here, by investigating the dynamics of these processes in the marine bacterium Hyphomonas neptunium, the authors unravel a two-step chromosomal segregation process reminiscent of eukaryotic mitosis, providing insights into the evolution of bacterial cell organization.
- Alexandra Jung
- , Anne Raßbach
- & Martin Thanbichler
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Article
| Open AccessFourier ring correlation simplifies image restoration in fluorescence microscopy
Fourier ring correlation (FRC) analysis is commonly used in fluorescence microscopy to measure effective image resolution. Here, the authors demonstrate that FRC can also be leveraged in blind image restoration methods, such as image deconvolution.
- Sami Koho
- , Giorgio Tortarolo
- & Giuseppe Vicidomini
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Article
| Open AccessThe cell-wide web coordinates cellular processes by directing site-specific Ca2+ flux across cytoplasmic nanocourses
Although calcium signals are known to be critical for many cellular processes, how signaling elicits specific functions remains unclear. In visually striking work, Duan et al. reveal that networks of cytoplasmic nanocourses orchestrate cell activity by directing site-specific calcium signals.
- Jingxian Duan
- , Jorge Navarro-Dorado
- & A. Mark Evans
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| Open AccessAnchoring cortical granules in the cortex ensures trafficking to the plasma membrane for post-fertilization exocytosis
Mammalian eggs require a single sperm for viable fertilization, and cortical granule exocytosis prevents additional sperm binding. Vogt et al. image at single granule resolution to document that cortical granule anchoring in the cortex ensures proper trafficking, exocytosis and polyspermy block.
- Edgar-John Vogt
- , Keizo Tokuhiro
- & Jurrien Dean
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| Open AccessComputational geometry analysis of dendritic spines by structured illumination microscopy
We are currently short of methods that can extract objective parameters of dendritic spines useful for their categorization. Authors present in this study an automatic analytical pipeline for spine geometry using 3D-structured illumination microscopy, which can effectively extract many geometrical parameters of dendritic spines without bias and automatically categorize spine population based on their morphological features
- Yutaro Kashiwagi
- , Takahito Higashi
- & Shigeo Okabe
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Article
| Open AccessPhotoregulated fluxional fluorophores for live-cell super-resolution microscopy with no apparent photobleaching
Super-resolution microscopy with spontaneously blinking dyes is dependent on pH and polarity of the medium. Here the authors introduce a photoactivatable fluxional fluorophore for live cell imaging that allows control over the fraction of spontaneously blinking molecules independently of medium properties.
- Elias A. Halabi
- , Dorothea Pinotsi
- & Pablo Rivera-Fuentes
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Article
| Open AccessMemory and relatedness of transcriptional activity in mammalian cell lineages
Phenotypically identical mammalian cells often display considerable variability in transcript levels of individual genes. Here the authors document how different genes propagate their expression levels in cell lineages and suggest a potential role of transcriptional memory for generating spatial patterns of gene expression.
- Nicholas E. Phillips
- , Aleksandra Mandic
- & David M. Suter
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Article
| Open AccessLive imaging of alveologenesis in precision-cut lung slices reveals dynamic epithelial cell behaviour
The process of alveologenesis is incompletely understood, partly due to the lack of applicable real-time imaging methods. Here the authors describe the process of alveologenesis and the behaviour of epithelial cells in real-time, using widefield microscopy and image deconvolution in precision-cut lung slices, revealing the dominant role of epithelial cell migration.
- Khondoker M. Akram
- , Laura L. Yates
- & Charlotte H. Dean
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Article
| Open AccessStem cell proliferation is induced by apoptotic bodies from dying cells during epithelial tissue maintenance
Damaged epithelial tissues are known to compensate for cell death through compensatory cell divisions to maintain epithelial integrity. Here, the authors show in living epithelia that dying cells stimulate adjacent stem cells to divide through caspase-dependent production of Wnt8a-containing apoptotic bodies.
- Courtney K. Brock
- , Stephen T. Wallin
- & George T. Eisenhoffer
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Article
| Open AccessQuantitative analysis of plant ER architecture and dynamics
Quantitative study of endoplasmic reticulum (ER) structure and dynamics has been a challenge. Here, the authors introduce software to automatically extract ER network elements from multi-dimensional fluorescence images of plant ER and to quantify structure, topology, protein localization and dynamics.
- Charlotte Pain
- , Verena Kriechbaumer
- & Mark Fricker
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Article
| Open AccessSecreted metalloproteases ADAMTS9 and ADAMTS20 have a non-canonical role in ciliary vesicle growth during ciliogenesis
Ciliogenesis is a complex process requiring hundreds of molecules, although few secreted proteins have been implicated. Here, the authors show that the secreted metalloproteases ADAMTS9 and ADAMTS20 intracellularly regulate ciliogenesis from unique periciliary vesicles with proteolytic activity.
- Sumeda Nandadasa
- , Caroline M. Kraft
- & Suneel S. Apte
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Article
| Open AccessLabel-free spatio-temporal monitoring of cytosolic mass, osmolarity, and volume in living cells
Label-free, spatio-temporal imaging of cellular physiological responses is challenging. Here the authors combine digital holographic microscopy with a millifluidic chip and mathematical modelling to quantify cell volume, mass and cell uptake under changing environmental conditions.
- Daniel Midtvedt
- , Erik Olsén
- & Gavin D. M. Jeffries
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Article
| Open AccessA computational framework to study sub-cellular RNA localization
Automated analysis of RNA localisation in smFISH data has been elusive. Here, the authors simulate and use a large dataset of images to design and validate a framework for highly accurate classification of sub-cellular RNA localisation patterns from smFISH experiments.
- Aubin Samacoits
- , Racha Chouaib
- & Florian Mueller
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Article
| Open AccessAXER is an ATP/ADP exchanger in the membrane of the endoplasmic reticulum
Endoplasmic reticulum (ER) functioning requires a constant supply of ATP, but the exchanger required for ATP uptake into the ER is unknown. Here, the authors report that SLC35B1, here named AXER, or ATP/ADP exchanger into the ER, can transport ATP into the ER.
- Marie-Christine Klein
- , Katharina Zimmermann
- & Ilka Haferkamp
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Article
| Open AccessVertebrate myosin 1d regulates left–right organizer morphogenesis and laterality
The actin-based motor Myosin1d is needed to establish left–right asymmetry in Drosophila. Here the authors show that myosin 1d has a role in lumen formation, vacuole trafficking and left-right asymmetry establishment during zebrafish development.
- Manush Saydmohammed
- , Hisato Yagi
- & Michael Tsang
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Article
| Open AccessConditional control of fluorescent protein degradation by an auxin-dependent nanobody
Current approaches to conditionally deplete target proteins require site-specific genetic engineering or have poor temporal control. Here the authors overcome these limitations by combining the AID system with nanobodies to reversibly degrade GFP-tagged proteins in living cells and zebrafish.
- Katrin Daniel
- , Jaroslav Icha
- & Jörg Mansfeld
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Article
| Open AccessAND-1 fork protection function prevents fork resection and is essential for proliferation
AND-1, the vertebrate orthologue of Ctf4, is a critical player during DNA replication and for maintenance of genome integrity. Here the authors use a conditional AND-1 depletion system in avian DT40 cells to reveal the consequences of the lack of AND-1 on cell proliferation and DNA replication.
- Takuya Abe
- , Ryotaro Kawasumi
- & Dana Branzei
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Article
| Open AccessA stochastic assembly model for Nipah virus revealed by super-resolution microscopy
The current model for Nipah virus assembly suggests that packaging is orchestrated by the matrix protein at the plasma membrane. Here, using super-resolution microscopy and Nipah virus-like particles, Liu et al. show that Nipah virus assembly is stochastic.
- Qian Liu
- , Lei Chen
- & Keng C. Chou
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Article
| Open AccessConcerted ESCRT and clathrin recruitment waves define the timing and morphology of intraluminal vesicle formation
Intraluminal vesicles are formed by the endosomal sorting complex required for transport (ESCRT) machinery. Here, the authors unravel the timing of vesicle budding, and that endosomal clathrin regulates concerted recruitment of ESCRT subcomplexes, required for efficient membrane remodeling.
- Eva Maria Wenzel
- , Sebastian Wolfgang Schultz
- & Camilla Raiborg