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. 2023 Nov 3;14:7071. doi: 10.1038/s41467-023-42820-3

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© The Author(s) 2023

Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — A Schematic of POI-eDHFR fusion construct. POI was directly fused to the N-terminus of eDHFR to allow for regulation with 7c. FLAG-tag was cloned downstream of eDHFR (to its C-terminus) for easy detection with anti-FLAG antibody. POI represents Lck, CD122, and RUNX1. B HEK293T-POI-eDHFR cells were incubated with 7c for 24 h. Western blot analysis was performed to probe for the expression of CD122, n = 3, C Lck, n = 3, and D RUNX1, n = 3, all detected with anti-FLAG antibody. CD122-eDHFR-FLAG fusion protein is 81 kDa, Lck-eDHFR-FLAG is 77 kDa, and RUNX1-eDHFR-FLAG is 73 kDa. COX IV = loading control, 18 kDa. E Immunofluorescence of cells expressing POI-eDHFR-FLAG constructs with AlexaFluor-488 secondary antibody against Anti-FLAG primary antibody (green), and DAPI nuclear staining (blue). From left to right: I. CD122-eDHFR-FLAG localized in the secretory pathway, II. LCK-eDHFR-FLAG membrane associated, and III. RUNX1-eDHFR-FLAG is in the nucleus. n = 3.