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A Cell-based Assay to Investigate Non-muscle Myosin II Contractility via the Folded-gastrulation Signaling Pathway in Drosophila S2R+ Cells
JoVE Journal
Biologia dello sviluppo
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JoVE Journal Biologia dello sviluppo
A Cell-based Assay to Investigate Non-muscle Myosin II Contractility via the Folded-gastrulation Signaling Pathway in Drosophila S2R+ Cells

A Cell-based Assay to Investigate Non-muscle Myosin II Contractility via the Folded-gastrulation Signaling Pathway in Drosophila S2R+ Cells

DOI:
10.3791/58325-v

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07:15 min

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August 19, 2018

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, , , , , , ,
1Department of Biology,The University of North Carolina at Chapel Hill, 2Department of Biology,Reed College, 3Carolina Center for Genome Sciences,The University of North Carolina at Chapel Hill, 4Lineberger Comprehensive Cancer Center,The University of North Carolina at Chapel Hill

Capitoli

  • 00:04Titolo
  • 01:10Preparation of Concanavalin-A-coated Glass-bottomed Dishes and the Plating of S2R+ Cells
  • 02:25Efficacy Test of the Fog-conditioned Medium and the Cellular Contractility Assay
  • 03:16Fixing and Staining of Constricted S2R+ Cells
  • 05:18Imaging and Quantification of the Cellular Contractility Assay
  • 05:41Results: Post-treatment with Fog-Myc, RLC Forms Rings Indicative of Non-Muscle Myosin II Constriction
  • 06:40Conclusion

Summary

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Here we describe a contractility assay using Drosophila S2R+ cells. The application of an exogenous ligand, folded gastrulation (Fog), leads to the activation of the Fog signaling pathway and cellular contractility. This assay can be used to investigate the regulation of cellular contractility proteins in the Fog signaling pathway.

Tags

Keywords: Non-muscle Myosin IIContractilityFolded-gastrulation Signaling PathwayDrosophila S2R+ CellsCell-based AssayCell DensityCell MorphologyCell Shape ChangeMorphogenesisDevelopmentGene DiscoveryRNAi Depletion

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