Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024:2795:105-111.
doi: 10.1007/978-1-0716-3814-9_11.

Phase Separation of Phytochrome B in HEK293T Cells

Di Chen  1   2 , Shangwei Zhong  3   4
Affiliations

Phase Separation of Phytochrome B in HEK293T Cells

Di Chen et al. Methods Mol Biol. 2024.

Abstract

In this method, we employed HEK293T cells to express the plant photoreceptor phytochrome B (phyB). Through the application of various treatments such as phycocyanobilin (PCB) supplementation, red light exposure, and temperature adjustments, the phyB proteins exhibited liquid-liquid phase separation, leading to the formation of biomolecular condensates. Here, we present a comprehensive description of the protein expression, cell treatment, and imaging capture procedures. This detailed guide provides step-by-step instructions on how to induce phase separation of phyB proteins in HEK293T cells. By utilizing this approach, researchers can investigate the physicochemical characteristics and dynamic formation process of phyB photobodies with precision.

Keywords: Biomolecular condensates; Light signal; Phase separation; Photobody; Phytochrome B; Temperature sensation.

PubMed Disclaimer

Similar articles

See all similar articles

References

    1. Quail PH, Boylan MT, Parks BM, Short TW, Xu Y, Wagner D (1995) Phytochromes: photosensory perception and signal transduction. Science 268(5211):675–680. https://doi.org/10.1126/science.7732376 - DOI - PubMed
    1. Sakamoto K, Nagatani A (1996) Nuclear localization activity of phytochrome B. Plant J 10(5):859–868. https://doi.org/10.1046/j.1365-313x.1996.10050859.x - DOI - PubMed
    1. Kircher L, Kozma-Bognar L, Kim L, Adam E, Harter K, Schafer E, Nagy F (1999) Light quality-dependent nuclear import of the plant photoreceptors phytochrome A and B. Plant Cell 11(8):1445–1456. https://doi.org/10.1105/tpc.11.8.1445 - DOI - PubMed - PMC
    1. Zhao Y, Shi H, Pan Y, Lyu M, Yang Z, Kou X, Deng XW, Zhong S (2023) Sensory circuitry controls cytosolic calcium-mediated phytochrome B phototransduction. Cell 186(6):1230–1243. https://doi.org/10.1016/j.cell.2023.02.011 - DOI - PubMed
    1. Van Buskirk EK, Reddy AK, Nagatani A, Chen M (2014) Photobody localization of Phytochrome B is tightly correlated with prolonged and light-dependent inhibition of hypocotyl elongation in the dark. Plant Physiol 165(2):595–560. https://doi.org/10.1104/pp.114.236661 - DOI - PubMed - PMC

LinkOut - more resources