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. 2023 Jun 7;11:1045759. doi: 10.3389/fcell.2023.1045759

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Copyright © 2023 Zhao, Veeranan-Karmegam, Baker, Mysona, Bagchi, Liu, Smith, Gonsalvez and Bollinger.

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) and the copyright owner(s) 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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Generation and validation of S1R-Apex cells. (A) Schematic of the proximity biotinylation strategy (B) HeLa cells stably expressing S1R-Apex were fixed and processed for immunofluorescence using a V5 antibody (green). S1R-Apex localizes around the nuclear envelope and to ER tubules. (C) HeLa cells stably expressing S1R-Apex were fixed and processed for immunofluorescence using a V5 antibody (green). The cells were also incubated with Streptavidin-647 (red) to reveal the localization of biotinylated proteins and were counterstained using DAPI (cyan). Biotinylated proteins co-localize with S1R-Apex. (D) Control HeLa cells not expressing a biotin ligase were processed using Streptavidin-647 (red). The cells were counterstained with DAPI (cyan). Minimal Streptavidin signal is observed in these cells. The scale bar in B is 15 microns and the scale bar in C is 20 microns.