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Dev Growth Differ 2022 Aug 01;646:306-317. doi: 10.1111/dgd.12794.
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Component analysis of nucleolar protein compartments using Xenopus laevis oocytes.

Lavering ED , Petros IN , Weeks DL .

The nucleolus is a multi-compartment, non-membrane-bound organelle within the nucleus. Nucleolar assembly is influenced by proteins capable of phase separation. Xenopus laevis oocytes contain hundreds of large nucleoli that provide experimental access for nucleoli that is unavailable in other systems. Here we detail methods to streamline the in vivo analysis of the compartmentalization of nucleolar proteins that are suspected of phase separation. The nucleolus is the main hub of ribosome biogenesis and here we present data supporting the division of proteins into nucleolar domains based on their function in ribosome biogenesis. We also describe the use of vital dyes such as Hoechst 33342 and Thioflavin T in nucleolar staining. Additionally, we quantify nucleolar morphology changes induced by heat shock and actinomycin D treatments. We suggest these approaches will be valuable in a variety of studies that seek to better understand the nucleolus, particularly those regarding phase separation. These approaches may also be instructive for other studies on phase separation, especially in the nucleus.

PubMed ID: 35607824
PMC ID: PMC9474603
Article link: Dev Growth Differ
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: elavl2 fbl gar1 gtpbp4 ncl nhp2 nop56 npm1 pak1ip1 pes1 rpl12 snu13

Article Images: [+] show captions
References [+] :
Alberti, Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates. 2019, Pubmed