Chapitre D'ouvrage Année : 2024

Analysis of Phase Separation of EARLY FLOWERING 3

Résumé

Phase separation is an important mechanism for regulating various cellular functions. The EARLY FLOWERING 3 (ELF3) protein, an essential element of the EVENING COMPLEX (EC) involved in circadian clock regulation, has been shown to undergo phase separation. ELF3 is known to significantly influence elongation growth and flowering time regulation and this is postulated to be due to whether the protein is in the dilute or phase separated state. Here, we present a brief overview of methods for analyzing ELF3 phase separation in vitro, including the generation of phase diagrams as a function of pH and salt versus protein concentrations, optical microscopy, Fluorescence Recovery After Photobleaching (FRAP) and turbidity assays.

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hal-04732679 , version 1 (11-10-2024)

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Maolin Peng, Stephanie Hutin, Aleksandra Mironova, Chloe Zubieta, Philip Wigge. Analysis of Phase Separation of EARLY FLOWERING 3. Meng Chen. Thermomorphogenesis : Methods and Protocols, 2795, Springer US, pp.123-134, 2024, Methods in Molecular Biology, ⟨10.1007/978-1-0716-3814-9_13⟩. ⟨hal-04732679⟩
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