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. 2014 May 16;5:203. doi: 10.3389/fpls.2014.00203

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Copyright © 2014 Hardré, Kuhn, Albrieux, Jouhet, Michaud, Seigneurin-Berny, Falconet, Block and Maréchal.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Chloroplast envelope tag and shave. Isolated chloroplasts were either untreated (control), treated with the non-permeable thermolysin protease (shave) or superficially biotinylated (tag), prior to envelope fractionation. Envelope proteins were subsequently resolved by 2D-PAGE as shown in Figure 2. Proteins were either stained with Coomassie blue (A,C) or transferred to a nitrocellulose membrane for Western-blotting using a specific markers of the outer envelope membrane, anti-OEP24 (B) or for biotin detection (D). A magnified detail of the gel is shown. Spot 73 that corresponds to OEP24 (black arrow) is detected with the antibody raised against OEP24 (B), and is consistently degraded (shaved) by the thermolysin protease (C) and biotinylated (tagged) by the biotin-XX,SE treatment. As a control, spot 69 (white arrow) is neither shaved nor tagged.