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. 2023 Jul 27;14(7):475. doi: 10.1038/s41419-023-06008-3

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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 Representative images of THOC3 localization in H520/H1703/SK-MES-1 cells detected by Fluorescent In Situ Hybridization (FISH) (40× magnification), scale bars = 100 μm (left); THOC3 mRNA expression in the nucleus and cytoplasm was determined via qRT-PCR in H1703/SK-MES-1/H520 cells (right). B Silver staining of immunoprecipitation of Flag and IgG antibodies in H1703 cells. C Bar chart showing proteins with unused scores of Flag over 10, identified via MS analysis of H1703 cells following immunoprecipitation with Flag and IgG antibodies (top); representative signaling pathways composed of proteins bound to THOC3 based on GO analysis (bottom). D THOC3 expression in cells transfected with si-NC or si-CCT8/CCT6 shown by WB. E Immunoprecipitation of cell lysates from H1703 cells transfected with Flag-tagged THOC3 using anti-CCT8, anti-CCT6A, anti-Flag, or IgG antibody, and immunoprecipitates were blotted with anti-Flag/CCT8/CCT6A antibodies. F Immunofluorescence images showing the distribution of THOC3 (green) and CCT8/CCT6A (red) in H1703 cells. Scale bars = 100 μm. G Cells are transfected with Flag-tagged THOC3 for 18 h and then incubated with and without 50 μg/ml cycloheximide (CHX) for 5 h. Immunoprecipitates were blotted with anti-CCT8/CCT6A antibodies. H Cells expressing Flag tagged THOC3 are prepared in the presence or absence of ATP and MgCl₂ and analyzed via immunoprecipitation and WB by anti-Flag/CCT8/CCT6A antibodies.