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. 2017 Sep 25;8:1646. doi: 10.3389/fpls.2017.01646

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Copyright © 2017 Sheshukova, Komarova, Pozdyshev, Ershova, Shindyapina, Tashlitsky, Sheval and Dorokhov.

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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NbAELP of agroinjected leaves was revealed mainly in the cell wall fraction. (A) Western blot analysis with NbAELP-specific antibodies of subcellular fractions of N. benthamiana leaves at 3 day after agroinjection with A. tumefaciens containing “empty” vector (pBin19). AEP, porcine kidney mutarotase “Calzyme” (~400 ng). CW, cell wall fraction; P1, fraction enriched with nuclear proteins; P30, fraction containing mainly membrane proteins from Golgi apparatus and endoplasmic reticulum. (B,C) Western blot analysis using anti-FLAG antibodies of N. benthamiana leaf proteins at 3 days after agroinjection with the 35S-NbAELP:3xFLAG vector. Cell homogenate was fractionated into CW, P1, P30, S30 (fraction of soluble cytoplasmic proteins) (B) and apoplast (C) fractions. The lower panels show the protein loading control stained with Amido Black.