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. 2024 Aug 27;43(20):4699–4719. doi: 10.1038/s44318-024-00211-4

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© The Author(s) 2024

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 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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) The interaction of endogenous STIC2 and ribosomes was tested by sucrose density gradient centrifugation of leaf extracts of A. thaliana WT plants. The gradient fractions were separated by SDS-PAGE and applied to immunoblotting using the indicated antibodies. A representative experiment from two independent biological and two technical replicates is shown. (B) Isolated chloroplasts from A. thaliana WT were lysed and separated into stromal and thylakoid fractions by centrifugation. Samples equivalent to 5 µg chlorophyll of chloroplast extract (C), washed thylakoids (T), and stroma (S) were separated on SDS-PAGE and blotted for immunodetection using antisera raised against STIC2, Alb3 and the small subunit of Rubisco (SSU). The Alb3 insertase and SSU were used as controls for successful fractionation. A Coomassie blue stained SDS gel served as loading control. A representative experiment from two independent biological replicates is shown. (C) Ribosome footprint yield was determined for soluble (yellow) and membrane (green) fractions of WT, stic2-3, ffc1-2, and ffc1-2 stic2-3 and normalized to the amount of fresh weight used as starting material. Means and SDs are derived from two (ffc1-2) or three (WT, stic2-3, ffc1-2 stic2-3) independent biological replicates. Source data are available online for this figure.