ArticleMethods in cell biology2017
Xenopus extract approaches to studying microtubule organization and signaling in cytokinesis.
Article in Methods in cell biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 44 citations in OpenAlex.
- Microtubule posttranslational modifications provide unique recognition patterns for associated proteins.The EMBO journal · 2026Article
- Glycogen-dependent demixing of frog egg cytoplasm at increased crowding.Biophysical journal · 2026Article
- An acentrosomal aster with atypical microtubule polarity recruits cytokinesis signals to its center in Xenopus egg extracts.Journal of cell science · 2025Article
- Episodic transport of protein aggregates achieves a positive size selectivity in aggresome formation.Nature communications · 2025Article
- Super-Resolution Mapping and Quantification of Molecular Diffusion via Single-Molecule Displacement/Diffusivity Mapping (SMAccounts of chemical research · 2025Article
- Density-dependent flow generation in active cytoskeletal fluids.Scientific reports · 2024Article
- Single-molecule diffusivity quantification inProceedings of the National Academy of Sciences of the United States of America · 2024Article
- Nuclear growth and import can be uncoupled.Molecular biology of the cell · 2024Article
- Branched microtubule nucleation and dynein transport organize RanGTP asters inMolecular biology of the cell · 2024Article
- Dorsal lip maturation and initial archenteron extension depend on Wnt11 family ligands.Developmental biology · 2023Article
- Dynein-dependent collection of membranes defines the architecture and position of microtubule asters in isolated, geometrically confined volumes of cell-free extracts.Molecular biology of the cell · 2022Article
- Nuclear F-actin and Lamin A antagonistically modulate nuclear shape.Journal of cell science · 2022Article
- Lysate-based pipeline to characterize microtubule-associated proteins uncovers unique microtubule behaviours.Nature cell biology · 2022Article
- Using FLIM-FRET for Characterizing Spatial Interactions in the Spindle.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Rho and F-actin self-organize within an artificial cell cortex.Current biology : CB · 2021Article
- Reconstitution of muscle cell microtubule organization in vitro.Cytoskeleton (Hoboken, N.J.) · 2021Article
- Xenopus laevis Egg Extract Preparation and Live Imaging Methods for Visualizing Dynamic Cytoplasmic Organization.Journal of visualized experiments : JoVE · 2021Article
- Spatial variation of microtubule depolymerization in large asters.Molecular biology of the cell · 2021Article
- Affinity Purification of Label-free Tubulins fromSTAR protocols · 2020Article
- Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
We report optimized methods for preparing actin-intact Xenopus egg extract. This extract is minimally perturbed, undiluted egg cytoplasm where the cell cycle can be experimentally controlled. It contains abundant organelles and glycogen and supports active metabolism and cytoskeletal dynamics that closely mimic egg physiology. The concentration of the most abundant ∼11,000 proteins is known from mass spectrometry. Actin-intact egg extract can be used for analysis of actin dynamics and interaction of actin with other cytoplasmic systems, as well as microtubule organization. It can be spread as thin layers and naturally depletes oxygen though mitochondrial metabolism, which makes it ideal for fluorescence imaging. When combined with artificial lipid bilayers, it allows reconstitution and analysis of the spatially controlled signaling that positions the cleavage furrow during early cytokinesis. Actin-intact extract is generally useful for probing the biochemistry and biophysics of the large Xenopus egg. Protocols are provided for preparation of actin-intact egg extract, control of the cell cycle, fluorescent probes for cytoskeleton and cytoskeleton-dependent signaling, preparation of glass surfaces for imaging experiments, and immunodepletion to probe the role of specific proteins and protein complexes. We also describe methods for adding supported lipid bilayers to mimic the plasma membrane and for confining in microfluidic droplets to explore size scaling issues.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.