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. 2023 Feb 18;16:8. doi: 10.1186/s13072-023-00481-y

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© The Author(s) 2023

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — TurboID-tagged KDM5 proteins are functional and biotinylate endogenous proteins. A Schematic of the workflow used for purifying and fingerprinting biotinylated proteins from adult heads using LC–MS/MS mass spectrometry. B Schematic of the four UASp constructs generated able to express HA-tagged TurboID alone, dCas9:TurboID, NT-KDM5 and CT-KDM5. C Western blot using adult heads showing levels of expression of KDM5 using anti-HA, biotinylation using a streptavidin-680 conjugate and the loading control alpha-tubulin. Genotypes: kdm5¹⁴⁰; gkdm5:HA (a wild-type strain; Control), Ubi-Gal4/+ ; UASp-TurboID/+ (TurboID), Ubi-Gal4/+ ; UASp-dCas9:TurboID (dCas9:TurboID), kdm5¹⁴⁰, Ubi-Gal4/kdm5¹⁴⁰; UASp-NT- kdm5/+ (NT-KDM5) and kdm5¹⁴⁰, Ubi-Gal4/kdm5¹⁴⁰; UASp-CT- kdm5 /+ (CT-KDM5). D Rescue of kdm5¹⁴⁰-induced lethality by ubiquitous expression of UAS-NT-kdm5 or UAS-CT-kdm5 using Ubi-Gal4. Genotype of male and female adult flies shown is kdm5¹⁴⁰, Ubi-Gal4/kdm5¹⁴⁰; UASp-NT- kdm5 /+ (NT-KDM5) and kdm5¹⁴⁰, Ubi-Gal4/kdm5.¹⁴⁰; UASp-CT- kdm5/+ (CT-KDM5)