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- W2072232356 abstract "The contractile actin cortex drives many cellular processes, such as cell migration and cytokinesis, but little is known about the proteins that regulate its assembly. To address this question, we developed an assay for assessing the involvement of candidate proteins in the recruitment of the main cortical components, actin and myosin, during cortex build-up. One situation where cortex assembly can easily be studied is during the growth of blebs. Blebs are membrane protrusions that are initially devoid of cortical proteins and that subsequently reassemble a cortical layer prior to bleb retraction. They therefore constitute an ideal system for the study of de novo cortex assembly under physiological conditions. In the developed assay, we use laser ablation of the cell cortex to induce bleb growth in a controlled manner and subsequently quantitatively monitor the recruitment of fluorescently labelled actin and myosin during the bleb life cycle. This allows for the extraction of a range of dynamic parameters of cortical assembly that can be compared between control cells and cells with varying levels of candidate proteins. Preliminary data, obtained with this assay, show that proteins typically involved in actin polymerization, such as profilin and cofilin, influence the rates of cortex assembly in HeLa cells. Additionally, the assay allowed us to precisely characterize the dynamics of cortex assembly in control cells, providing new insights into the mechanisms of bleb growth and retraction. From these first tests, we conclude that the developed assay provides a highly sensitive tool for the study of cortex assembly." @default.
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- W2072232356 date "2010-01-01" @default.
- W2072232356 modified "2023-10-16" @default.
- W2072232356 title "Analysis of De Novo Cell Cortex Assembly in Blebs as a Novel Assay for Probing Cortical Dynamics and Regulation" @default.
- W2072232356 doi "https://doi.org/10.1016/j.bpj.2009.12.853" @default.
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