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. 2020 Feb 26;18:19. doi: 10.1186/s12915-020-0749-y

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© The Author(s). 2020

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 7 — The Sec61 translocon polypeptide channel does not participate in glutathione transport. Effects of manipulating the translocon on fluorescence ratio changes of Grx1-roGFP1-iE_ER in HEK293 cells stably expressing the sensor. Each trace corresponds to the data recorded from one cell. At the end of each experiment, 500 μM diamide (Dia) and 20 mM DTT were added to ensure the functionality of the probe. a One hundred micromolar puromycin, b 200 μM anisomycin followed by 100 μM puromycin, e 200 μM anisomycin followed by 1 μM TG, and f 100 μM puromycin followed by 1 μM TG were applied as indicated by the arrow. c Cells were treated overnight with 1 mM BSO prior to experiment, and 100 μM puromycin were applied as marked. d HEK293 cells stably expressing Grx1-roGFP1-iE_ER were transfected with control or Sec61 siRNA for 48 h before imaging as above; 1 μM TG were applied as indicated by the arrow. Knockdown efficiency was verified by Western blot (aSec61a, anti-Sec61α antibody; aBiP, anti-BiP antibody; aActin, anti-actin antibody)