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Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc
Apicobasal polarity determinants are potential tumor suppressors that have been extensively studied. However, the precise mechanisms by which their misregulation disrupts tissue homeostasis are not fully understood. Here, we present a comprehensive protocol for establishing a conditional RNAi knockd...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10550844/ https://www.ncbi.nlm.nih.gov/pubmed/37768826 http://dx.doi.org/10.1016/j.xpro.2023.102566 |
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author | Antson, Hanna Huang, Yunxian Tõnissoo, Tambet Shimmi, Osamu |
author_facet | Antson, Hanna Huang, Yunxian Tõnissoo, Tambet Shimmi, Osamu |
author_sort | Antson, Hanna |
collection | PubMed |
description | Apicobasal polarity determinants are potential tumor suppressors that have been extensively studied. However, the precise mechanisms by which their misregulation disrupts tissue homeostasis are not fully understood. Here, we present a comprehensive protocol for establishing a conditional RNAi knockdown of scribble in Drosophila wing imaginal disc. We describe steps for generating fly lines, conditional knockdown in host stocks, and sample preparation. We then detail procedures for imaging, image analysis, and verification of wing disc phenotypes by various antibodies. For complete details on the use and execution of this protocol, please refer to Huang et al.(1) |
format | Online Article Text |
id | pubmed-10550844 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-105508442023-10-06 Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc Antson, Hanna Huang, Yunxian Tõnissoo, Tambet Shimmi, Osamu STAR Protoc Protocol Apicobasal polarity determinants are potential tumor suppressors that have been extensively studied. However, the precise mechanisms by which their misregulation disrupts tissue homeostasis are not fully understood. Here, we present a comprehensive protocol for establishing a conditional RNAi knockdown of scribble in Drosophila wing imaginal disc. We describe steps for generating fly lines, conditional knockdown in host stocks, and sample preparation. We then detail procedures for imaging, image analysis, and verification of wing disc phenotypes by various antibodies. For complete details on the use and execution of this protocol, please refer to Huang et al.(1) Elsevier 2023-09-28 /pmc/articles/PMC10550844/ /pubmed/37768826 http://dx.doi.org/10.1016/j.xpro.2023.102566 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Antson, Hanna Huang, Yunxian Tõnissoo, Tambet Shimmi, Osamu Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title | Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title_full | Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title_fullStr | Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title_full_unstemmed | Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title_short | Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc |
title_sort | conditional knockdown protocol for studying cellular communication using drosophila melanogaster wing imaginal disc |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10550844/ https://www.ncbi.nlm.nih.gov/pubmed/37768826 http://dx.doi.org/10.1016/j.xpro.2023.102566 |
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