Detection of changes in the Medicago sativa retinoblastoma-related protein (MsRBR1) phosphorylation during cell cycle progression in synchronized cell suspension culture

Ferhan Ayaydin, Edit Kotogány, Edit Ábrahám, Gábor V. Horváth

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Deepening our knowledge on the regulation of the plant cell division cycle depends on techniques that allow for the enrichment of cell populations in defined cell cycle phases. Synchronization of cell division can be achieved using different plant tissues; however, well-established cell suspension cultures provide large amount of biological sample for further analyses. Here, we describe the methodology of the establishment, propagation, and analysis of a Medicago sativa suspension culture that can be used for efficient synchronization of the cell division. A novel 5-ethynyl-2'-deoxyuridine (EdU)-based method is used for the estimation of cell fraction that enters DNA synthesis phase of the cell cycle and we also demonstrate the changes in the phosphorylation level of Medicago sativa retinoblastoma-related protein (MsRBR1) during cell cycle progression.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages267-285
Number of pages19
Volume1524
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1524
ISSN (Print)10643745

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Keywords

  • 5-Ethynyl-2ʹ-deoxyuridine staining
  • Cell cycle synchronization
  • Fluorescence microscopy
  • Hydroxyurea
  • Medicago sativa suspension culture
  • Retinoblastoma-related protein phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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