Photobody Detection Using Immunofluorescence and Super-Resolution Imaging in Arabidopsis

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2021 Mar 3

PMID 33656665

Citations 1

Authors

Giorgio Perrella

Anna Zioutopoulou

Andrew Hamilton

Eirini Kaiserli

Affiliations

Soon will be listed here.

Abstract

Light triggers changes in plant nuclear architecture to control differentiation, adaptation, and growth. A series of genetic, molecular, and imaging approaches have revealed that the nucleus forms a hub for photo-induced protein interactions and gene regulatory events. However, the mechanism and function of light-induced nuclear compartmentalization is still unclear. This chapter provides detailed experimental protocols for examining the morphology and potential functional significance of light signaling components that localize in light-induced subnuclear domains, also known as photobodies. We describe how immunolabeling of endogenous proteins and fluorescent in situ hybridization (FISH) could be combined with confocal imaging of fluorescently tagged proteins to assess co-localization in Arabidopsis nuclei. Furthermore, we employ a super-resolution imaging approach to study the morphology of photobodies at unprecedented detail.

Citing Articles

Out of the Dark and Into the Light: A New View of Phytochrome Photobodies.

Pardi S, Nusinow D Front Plant Sci. 2021; 12:732947.

PMID: 34531891 PMC: 8438518. DOI: 10.3389/fpls.2021.732947.

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